Structure and function of Full-length Tau

Laura Vallés-Saiz; Indalo Domene-Serrano; Angel J. Picher; Mar Pérez; Vega García-Escudero; Félix Hernández; Jesús Avila

doi:10.1371/journal.pone.0335251

Peer Review History

Original SubmissionFebruary 11, 2025
8 Aug 2025 Decision Letter - Stephen D. Ginsberg, Editor Dear Dr. Avila, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Sep 21 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Stephen D. Ginsberg, Ph.D. Section Editor PLOS ONE Journal Requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. We note that the grant information you provided in the ‘Funding Information’ and ‘Financial Disclosure’ sections do not match. When you resubmit, please ensure that you provide the correct grant numbers for the awards you received for your study in the ‘Funding Information’ section. 3. Thank you for stating the following financial disclosure: “This work has been supported by grants from the Spanish Ministry of Science: PID2020-113204GB-I00 (F.H.) and PID2021-123859OB-100 from MCIN /AEI/10.13039/501100011033 / FEDER, UE (J.A.). The Centro de Biología Molecular Severo Ochoa (CBMSO) is a Severo Ochoa Center of Excellence (MICIN, award CEX2021-001154-S).” Please state what role the funders took in the study. If the funders had no role, please state: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." If this statement is not correct you must amend it as needed. Please include this amended Role of Funder statement in your cover letter; we will change the online submission form on your behalf. 4. Please note that funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript. 5. When completing the data availability statement of the submission form, you indicated that you will make your data available on acceptance. We strongly recommend all authors decide on a data sharing plan before acceptance, as the process can be lengthy and hold up publication timelines. Please note that, though access restrictions are acceptable now, your entire data will need to be made freely accessible if your manuscript is accepted for publication. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If you are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process. 6. Thank you for stating the following in the Financial Disclosure section: “This work has been supported by grants from the Spanish Ministry of Science: PID2020-113204GB-I00 (F.H.) and PID2021-123859OB-100 from MCIN /AEI/10.13039/501100011033 / FEDER, UE (J.A.). The Centro de Biología Molecular Severo Ochoa (CBMSO) is a Severo Ochoa Center of Excellence (MICIN, award CEX2021-001154-S).” We note that one or more of the authors are employed by a commercial company: Expedeon SLU 1) Please provide an amended Funding Statement declaring this commercial affiliation, as well as a statement regarding the Role of Funders in your study. If the funding organization did not play a role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript and only provided financial support in the form of authors' salaries and/or research materials, please review your statements relating to the author contributions, and ensure you have specifically and accurately indicated the role(s) that these authors had in your study. You can update author roles in the Author Contributions section of the online submission form. Please also include the following statement within your amended Funding Statement. “The funder provided support in the form of salaries for authors [insert relevant initials], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.” If your commercial affiliation did play a role in your study, please state and explain this role within your updated Funding Statement. 2) Please also provide an updated Competing Interests Statement declaring this commercial affiliation along with any other relevant declarations relating to employment, consultancy, patents, products in development, or marketed products, etc. Within your Competing Interests Statement, please confirm that this commercial affiliation does not alter your adherence to all PLOS ONE policies on sharing data and materials by including the following statement: ""This does not alter our adherence to PLOS ONE policies on sharing data and materials.” (as detailed online in our guide for authors http://journals.plos.org/plosone/s/competing-interests) . If this adherence statement is not accurate and there are restrictions on sharing of data and/or materials, please state these. Please note that we cannot proceed with consideration of your article until this information has been declared. Please include both an updated Funding Statement and Competing Interests Statement in your cover letter. We will change the online submission form on your behalf. 7. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 8. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. 9. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? Reviewer #1: Partly Reviewer #2: Yes Reviewer #3: Yes Reviewer #4: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes Reviewer #4: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes Reviewer #4: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: Yes Reviewer #4: No ****** Reviewer #1: The manuscript analyzes, by structural and functional approaches, the isoform of Tau protein named the full tau isoform, containing all exons of the MAPT gene. The paper's content is of interest in the field; however, some implementations are required to be more understandable for the readers and Tau researchers. Furthermore, a key mechanism of action of the Tau protein or Tau isoforms, the phosphorylation of the protein, was not considered, in either the discussion or the experimental design. The material and methods are adequately described, and the results are clear. Major concern: The authors have produced the Tau full length utilizing a bacterial model. Since protein phosphorylation is an essential post-translational modification for its function and aggregation mechanism, did the authors verify whether lentiviruses' protein was phosphorylated? Could phosphorylation influence functional aggregation assays? The authors should comment this aspect in the discussion section. It would also be appropriate to verify if the protein is phosphorylated in expression studies on viability and growth. Authors might declare the limit of the study. Minor point: - The denomination tau 42 must be explained by the authors in the manuscript's text. - In the introduction (lines 66-70), it will be helpful to include at least an additional and recent reference that supports the statement (such as https://doi.org/10.1038/ s41580-022-00545- z or others). - Lines 64-65, the sentence (Where there are…) is unclear. - Lines 94-97 check the English style. - Line 238 "To try to explain the previous, we decide" seems that a world is missing; it might be the world "results". - To be more informative, specify in figure 1 and the figure caption the differences between panel B (protein bacterial purification) and panel C, western blot using Tau-specific antibody. In the discussion, authors might declare the limit of the study. Reviewer #2: In the article entitled “Structure and function of full-length Tau” the authors analyze the full-length Tau isoform by comparing it with Tau 42. There are a few points that are important to address. 1. Regarding biophysical characterization, aggregation and structure are shown by microscopy, however fluorescence spectroscopy or conformational dynamics data would be missing. 2. Modeling by AlphaFold2 is useful, but the conclusion on domain accessibility is not experimentally validated, such as limited proteolysis assays or FRET. 3. The presence of amorphous aggregates with Full Tau could be characterized by immuno-MS. 4. The amount of Tau in soluble fraction is not shown, which complements the analysis. 5. Regarding Figure 3, the images shown are qualitative, however, they could be improved with a quantitative analysis of the number, length or diameter of the aggregates. A morphological classification could be included, for example fibrillar vs. amorphous. 6. It would be necessary to examine aggregation in the absence of heparin. 7. It is not shown if the aggregates are beta structures. 8. It is not known whether the amorphous Full Tau aggregates are transiently toxic or inert. 9. Regarding figure 4, no quantitative value of surface accessibility or electrostatic maps are provided. 10. A comparative analysis of free energy, intramolecular interactions or contact prediction to support the model is missing. 11. Regarding Figure 5, the use of a single cell line limits the extrapolation to neuronal models. It would be useful to evaluate markers of apoptosis (TUNEL, caspases) to confirm the mechanism of cell death. Overall, this is a very interesting article that explores the complete Tau isoform, through expression, microscopy, cell functional analysis, modeling and statistical quantification. Reviewer #3: The manuscript is well written. It is a topic of great importance, and the authors have carefelly studied the role of exons in the structure and function of tau. 1. The grammar needs to be checked throughout the manuscript, as some sentences do not make sense. 2. In introduction, 'Three main Tau transcripts from the MAPT gene have been described", needs elaboration. 3. IN the same paragraph, "none have been found to translate the full-length tau protein" needs more citations and elaboration. It may help the readers if a few sentences on why it has not been possible, can be mentioned. 4. next paragraph of introduction, "essential roles in tau protein" - what kind of roles? 5. Abbreviatons like pLDDT need to be elaborated. 6. A conclusion discussing the significance of the findings may be included, as it will show the author's opinions and future perspectives. Reviewer #4: The manuscript addresses an interesting topic in the field of neuroscience: the first study of the structure and function of the complete Tau isoform in the brain. While research on the Tau protein has been a topic of study for years, the proposal presented by the authors provides relevant new information that makes the work a valuable contribution to its field. Overall, the manuscript is well-structured and written, allowing for guided reading and supported by robust evidence. The conclusions are appropriate for the study's objective and the scientific argument presented. Regarding the cited literature, fundamental studies and reviews recognized in the field are mentioned. One-sixth of the references are updated; in this case, this could be due to the limited literature on the topic. Minor revisions are suggested: - The image clarity is low. Increasing the figure resolution is recommended. - The legend for Figure 1 is incomplete; the abbreviations mentioned in the image do not indicate their meaning. - Correct the spelling of Control Mts to Control MTs (Figure 2B). - Add the meaning of the abbreviations used (SN, PLL) to the descriptions of Figures 2C and 3B. - Correct the spelling of the abbreviation Pll to PLL (Figure 3C). - The text mentions that the filaments in Figure 3 are marked with a white arrow, but this is not the case. - Latin words should be written in italics: post hoc. - Correctly spell the chemical formula MgCl2. - Write the chemical formula of ammonium acetate. - Based on the results obtained and the available literature, what do the authors believe could be the substantial function of the complete Tau protein in the brain? ****** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: No Reviewer #2: No Reviewer #3: No Reviewer #4: Yes: Donaji Chi-Castañeda ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step. Attachments Attachment Submitted filename: PONE-D-25-07400(1).pdf https://doi.org/10.1371/journal.pone.0335251.r002
Revision 1
15 Sep 2025 Author Response Dear Prof. Ginsberg, Editor Plos One. Please find enclosed the revised manuscript entitled “Structure and function of full-length Tau.” In this revised version, we have carefully addressed all the points raised by the editor and reviewers, as you indicated. Journal requirements: 1. We have ensured that our manuscript and figures comply with PLOS ONE’s style requirements, following the templates provided by the journal. 2. Grant numbers will be double-checked when resubmitting the manuscript. 3. The funders’ role has been added to the financial disclosure section. The amended text now reads: “This work has been supported by grants from the Spanish Ministry of Science: PID2023-149460NB-I00 (F.H.: Conceptualization, Formal analysis, Funding acquisition, Project Writing – original draft and Writing – review and editing) and PID2021-123859OB-100 from MCIN /AEI/10.13039/501100011033 / FEDER, UE (J.A.: Conceptualization, Formal analysis, Funding acquisition, Project Writing – original draft and Writing – review and editing). 4basebio S.L.U. supported AJP in the form of salary and research materials for the study. The Centro de Biología Molecular Severo Ochoa (CBMSO) is a Severo Ochoa Center of Excellence (MICIN, award CEX2021-001154-S).” 4. Funding information provided in other sections has been removed. 5. Data sharing will be indicated in the submission form. The data are available in the repository: https://doi.org/10.6084/m9.figshare.28351508.v1 6. Roles, responsibilities, and conflicts of interest have been updated to fully adhere to PLOS ONE’s policies and procedures. 7. Supporting information captions have been included at the end of the manuscript. 8. Raw, uncropped, and unadjusted images are included either in the Supporting Information or in the data repository. Reviewer 1: Major Concern The authors have produced the Tau full length utilizing a bacterial model. Since protein phosphorylation is an essential post-translational modification for its function and aggregation mechanism, did the authors verify whether lentiviruses' protein was phosphorylated? Could phosphorylation influence functional aggregation assays? The authors should comment this aspect in the discussion section. It would also be appropriate to verify if the protein is phosphorylated in expression studies on viability and growth. Full Tau and Tau 4R2N, originate from bacterial cloning and purification. Thus, both proteins lack of post-translational modifications like, phosphorylation, acetylation, glycosylation, … We have been working with unmodified proteins in this first manuscript to focus on those proteins without any modifications. Indeed, many works from other groups had been also performed with unmodified tau with reliable results. In future studies, we will analyze how different types of modifications at specific sites can affect to the function or disfunction of Full Tau isoform. Moreover, the aim of this study is to investigate the function and structure of the full-length Tau protein in the physiological context of the isoform, rather than in the pathological setting where its modifications would indeed occur. Minor points: 1. The denomination tau 42 must be explained by the authors in the manuscript's text. The term Tau 42 referred to the tau isoform Tau 4R2N. This has now been changed to the canonical denomination for clarity. 2. In the introduction (lines 66-70), it will be helpful to include at least an additional and recent reference that supports the statement (such as https://doi.org/10.1038/ s41580-022-00545- z or others). Several references have been added to the statement. 3. Lines 64-65, the sentence (Where there are…) is unclear. The sentence “Where there are…” has been rewritten for clarification. 4. Lines 94-97 check the English style. English style has been checked and improved. 5. Line 238 "To try to explain the previous, we decide" seems that a world is missing; it might be the world "results". Grammar and style throughout the manuscript have been revised and clarified. 6. To be more informative, specify in figure 1 and the figure caption the differences between panel B (protein bacterial purification) and panel C, western blot using Tau-specific antibody. The caption for Figure 1 has been expanded, including a statement clarifying the origin of both panels. Reviewer 2: 1. Regarding biophysical characterization, aggregation and structure are shown by microscopy, however fluorescence spectroscopy or conformational dynamics data would be missing. We don’t consider that such analysis is related to the focus of the present work. Nevertheless, in the near future we will deeply study conformational changes on Full Tau in different environments. Since fluorescence spectroscopy is based on the excitation of the intrinsic fluorescent properties of the molecules analyzed, we do not consider that the results obtained from these assays directly support the objectives of the present project. However, the use of techniques aimed at analyzing conformational changes has indeed been proposed for future studies, which will focus in greater detail on the structural features of the isoform and their post translational modifications. 2. Modeling by AlphaFold2 is useful, but the conclusion on domain accessibility is not experimentally validated, such as limited proteolysis assays or FRET. The structural modeling obtained through the machine learning program AlphaFold2 illustrates the conformations adopted by the protein depending on the presence or absence of specific exons, and, notably, highlights the conformational differences associated with exon 8. These in silico data provide a rationale for the differences observed in the results presented in Figure 2. In this context, we do not fully see the necessity of including proteolysis assays or FRET analyses, as they are not essential to the main objectives of the present work. Nevertheless, we greatly appreciate the reviewer’s suggestion, and the implementation of these biophysical approaches will indeed be considered in future studies, in which we intend to explore these aspects in greater depth. 3. The presence of amorphous aggregates with Full Tau could be characterized by immuno-MS. Immuno-MS of amorphous aggregates with Full Tau has been performed and included in supporting information as S4. 4. The amount of Tau in soluble fraction is not shown, which complements the analysis. Soluble fraction fraction of both isoforms has been included in Figure 2, as suggested by the reviewer. 5. Regarding Figure 3, the images shown are qualitative, however, they could be improved with a quantitative analysis of the number, length or diameter of the aggregates. A morphological classification could be included, for example fibrillar vs. amorphous. Figure 3 has been improved with several quantitative data to illustrate every detail of the polymers. 6. It would be necessary to examine aggregation in the absence of heparin. Aggregation of Full Tau and Tau 4R2N, in the absence of heparin, has been included in supporting information as S5. 7. It is not shown if the aggregates are beta structures. We cannot rule out the presence of β-structures in the aggregates observed in the aggregation assay performed with Full Tau. Indeed, Full Tau shows some of those structures with canonical isoform. When comparing regions of the tau protein such as exons 10 and 11, which are known to form β-structures, we find substantial sequence similarities with exon 8. This resemblance is now shown in S6. Suggesting that exon 8 could also contribute to β-structural arrangements, in addition to exons 10 and 11, within the aggregates. 8. It is not known whether the amorphous Full Tau aggregates are transiently toxic or inert. These aggregates do not appear to be toxic, as shown in Figure 5, where cell survival following transduction with the Full Tau protein was not reduced compared to the control. This stands in contrast to cells transduced with the 4R2N isoform, where a marked decrease in viability was observed 9. Regarding figure 4, no quantitative value of surface accessibility or electrostatic maps are provided. The Solvent Accessible Surface Area (SASA) data have been incorporated into the Supporting Information. 10. A comparative analysis of free energy, intramolecular interactions or contact prediction to support the model is missing. We appreciate the suggestion of techniques that would further enhance and increase the value of the study. However, the primary aim of this work is to provide a functional and structural characterization that introduces the first insights into this isoform. These biophysical and biological approaches will indeed be implemented in future studies, where we will explore these aspects in greater depth. 11. Regarding Figure 5, the use of a single cell line limits the extrapolation to neuronal models. It would be useful to evaluate markers of apoptosis (TUNEL, caspases) to confirm the mechanism of cell death. As previously mentioned, the aim of this work is to indicate, for the first time, that Full Tau isoform could be expressed and that this tau isoform shows different characteristics to those found for canonical CNS tau isoforms, been Tau 4R2N a good example of those canonical tau isoforms. In this way, cells transduced with Full Tau did not undergo cell death, whereas cells transduced with Tau 4R2N did, consistent with previous reports of Tau 4R2N-induced cytotoxicity. That cell death could occur by Regulate Cell Death (RCD) as apoptosis, pyroptosis, autophagy and ferroptosis… In further studies, we will study those possibilities by the methodology indicated by the reviewer. We deeply appreciate the indications and comments of the reviewer because after doing some of the experiments suggested, the work has improved thanks to that input, and we anticipate that their ideas will also be useful in future research. Finally, we thank such a positive final sentence about the work. Reviewer 3: 1. The grammar needs to be checked throughout the manuscript, as some sentences do not make sense. Grammar and style throughout the manuscript have been revised and clarified. 2. In introduction, 'Three main Tau transcripts from the MAPT gene have been described", needs elaboration. The sentence “Three main Tau transcripts…” has been checked and elaborated. 3. In the same paragraph, "none have been found to translate the full-length tau protein" needs more citations and elaboration. It may help the readers if a few sentences on why it has not been possible, can be mentioned. More citations have been added to support the statement “none have been found…,” and the sentence has been rephrased for clarity. 4. Next paragraph of introduction, "essential roles in tau protein" - what kind of roles? The roles of tau protein are included with appropriate citations. 5. Abbreviatons like pLDDT need to be elaborated. Every abbreviation found in the manuscript has been included in the abbreviation section. 6. A conclusion discussing the significance of the findings may be included, as it will show the author's opinions and future perspectives. The sentences highlighted by the reviewer have been corrected and further explained in greater detail. Reviewer 4: 1. The image clarity is low. Increasing the figure resolution is recommended. Figures included in the revised manuscript have been uploaded at 600 dpi resolution to improve clarity. 2. The legend for Figure 1 is incomplete; the abbreviations mentioned in the image do not indicate their meaning. The caption for Figure 1 has been completed, with the abbreviations been included in the figure and in abbreviations section. 3. Correct the spelling of Control Mts to Control MTs (Figure 2B). Mts has been checked and changed by MTs. 4. Add the meaning of the abbreviations used (SN, PLL) to the descriptions of Figures 2C and 3B. Both abbreviations (SN and PLL) have been included in the figure caption and abbreviations section. 5. Correct the spelling of the abbreviation Pll to PLL (Figure 3C). Sll has been corrected to SLL. 6. The text mentions that the filaments in Figure 3 are marked with a white arrow, but this is not the case. White arrows have been added to Figure 3. 7. Latin words should be written in italics: post hoc. Latin words have been formatted in italics. 8. Correctly spell the chemical formula MgCl2 and write the chemical formula of ammonium acetate. Chemical formulas, both MgCl2 and ammonium acetate (NH4CH3CO2) have been corrected and included. 9. Based on the results obtained and the available literature, what do the authors believe could be the substantial function of the complete Tau protein in the brain? The amount of Full Tau is much lower than that of CNS canonical isoforms. This difficult the search of its possible function. A possible way to analyze that possible function of Full Tau will be to confirm its presence in any animal model at a specific localization. That data would facilitate the knowledge of its putative function. We will further work on that. Yours sincerely, Jesús Avila https://doi.org/10.1371/journal.pone.0335251.r003
25 Sep 2025 Decision Letter - Stephen D. Ginsberg, Editor Dear Dr. Avila, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Nov 09 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Stephen D. Ginsberg, Ph.D. Section Editor PLOS ONE Journal Requirements: If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: All comments have been addressed Reviewer #2: All comments have been addressed Reviewer #4: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #1: Yes Reviewer #2: Yes Reviewer #4: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes Reviewer #4: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes Reviewer #4: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes Reviewer #4: No ****** Reviewer #1: (No Response) Reviewer #2: Thank you to the authors for their replies. With the changes made, the manuscript has improved substantially. Reviewer #4: Standardize the spelling of Tau throughout the manuscript. Sometimes it appears as "tau" and sometimes as "Tau." The same applies to "full tau," which sometimes appears as "Full Tau." On line 206, the reference 32 is in parentheses, not brackets. On line 527, subscript the number 2 in the CaCl2 formula. ****** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: No Reviewer #2: No Reviewer #4: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org https://doi.org/10.1371/journal.pone.0335251.r004
Revision 2
7 Oct 2025 Author Response In this revised version, we have carefully addressed all the points raised by the reviewers, as you indicated. Reviewer 1 and 2: We thank them for their positive comments. Reviewer 4: 1. Is the manuscript presented in an intelligible fashion and written in standard English? The manuscript has been revised to improve grammar and clarity We believe it is now written in standard, intelligible English. 2. Standardize the spelling of Tau throughout the manuscript. Sometimes it appears as "tau" and sometimes as "Tau." The same applies to "full tau," which sometimes appears as "Full Tau." The spelling of Tau and Full Tau has been standardized throughout the manuscript. 3. On line 206, the reference 32 is in parentheses, not brackets. The reference has been corrected to [32]. 4. On line 527, subscript the number 2 in the CaCl2 formula. The chemical formula has been corrected to show CaCl₂ with the subscript as indicated. We also thank the reviewer for these helpful comments. Attachments Attachment Submitted filename: Response to Editors.docx https://doi.org/10.1371/journal.pone.0335251.r005
8 Oct 2025 Decision Letter - Stephen D. Ginsberg, Editor Structure and function of Full-length Tau PONE-D-25-07400R2 Dear Dr. Avila, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support . If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Stephen D. Ginsberg, Ph.D. Section Editor PLOS ONE https://doi.org/10.1371/journal.pone.0335251.r006
Formally Accepted
Acceptance Letter - Stephen D. Ginsberg, Editor PONE-D-25-07400R2 PLOS ONE Dear Dr. Avila, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Stephen D. Ginsberg Section Editor PLOS ONE https://doi.org/10.1371/journal.pone.0335251.r007

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .