PONE-D-22-31627The Structure of Prevacuolar Compartments in Neurospora crassa as observed with Super-Resolution MicroscopyPLOS ONE

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Reviewers' comments:

Comments to the Author

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Reviewer #1: Yes

Reviewer #2: No

Reviewer #3: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: N/A

Reviewer #2: N/A

Reviewer #3: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

Reviewer #3: Yes

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5. Review Comments to the Author

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Reviewer #1: Bowman analyzed PVCs in Neurospora crassa in detail by super-resolution microscopy using Rab7 as a marker. The results revealed a pouch-like structure, which contains dynein and dynactin. The following points need to be further examined:

1. L237-241, if the relationship between dynein localization in PVC and microtubules is to be further investigated, microtubule polymerization inhibitors should be used to see what happens to the localization.

2. L289-290, an actual example should be provided how vesicles are entering and exiting the PVC.

Italicize: L30 Neurospora; L296,297 Aspergillus nidulans; L301 Phanerochaete

L87, 2012a].

L154, which image was overexposed shown in Panel A?

L207, dynein intermediate chain (DIC)

Reviewer #2: The author describes a detailed observation of prevacuolar compartments (PVCs) in the filamentous fungus Neurospora crassa. The fungus exhibits a larger size and dynamic structures of PVCs, and thus it is a good model to characterize the organellar features. This study employs super-resolution microscopy, which revealed that the PVCs have a pouch shape with vesicles and tubules. Importantly, dynein and dynactin, components of microtubule motor, were surrounded with PVCs.

However, the present study does not contain many new findings because similar observations were already conducted by the same author (Bowman et al., 2015. Characterization of a Novel Prevacuolar Compartment in Neurospora crassa. Eukaryot Cell 14, 1253–1263). The observations of components such as Rab7 and dynein look mostly similar to those in the previous publication, in which PVCs were extensively characterized. Although dynamic movement of PVCs is shown in the supplementary movie, figure pictures do not illustrate scientifically significant aspects.

Comments.

l. 49 Forgac, 2007 is not related to the localization in hyphae.

l. 56-57 Should Bowman et al., 2015 be cited for the result of CPY and PHO-8?

l. 164 In the picture of Fig. 2B, the size of small vesicles or tubules is not shown clearly.

l. 238 No results on microtubules are shown.

Reviewer #3: The manuscript titled “The Structure of Prevacuolar Compartments in Neurospora crassa as observed with Super-Resolution Microscopy” by Barry Bowman describes very new findings about the prevacuolar compartments (PVCs) of Neurospora crassa. The manuscript is technically sound providing deeper insight to the PVC that appears to be rapidly changing in their structure.

The manuscript may be accepted after minor revisions as suggested below.

1. This is a single author manuscript. Therefore, “we” may be replaced with “I” in the sentences like (i) “By using super-resolution microscopy we have achieved a better understanding of the structure of the PVCs (line 21-23);

(ii) “We hypothesize that…. (line 65)”; and

(iii) “We previously reported … (line 245)” etc.

2. The reference to the figures are not consistent in the manuscript. At some places it is in the main sentence and at some other places it is in the parentheses,

e. g.

line: 138: shown in figure 1..

line 151. The image in Fig 1

line 165: (Fig 2B)…

It is better to use a consistent style, preferably referring to the figures within the parentheses as in the line 165.

3. Line 283: In our previous work we observed a possible maturation process in PVCs…

Please provide supporting reference for this line.

4. Fig. 1: Kindly insert a single space in the scale, it should be 5 μm.

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Submitted filename: Comments December 11 2022.docx

The Structure of Prevacuolar Compartments in Neurospora crassa as observed with Super-Resolution Microscopy

PONE-D-22-31627R1

Dear Dr. Bowman,

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.

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Kind regards,

Praveen Rao Juvvadi, PhD

Academic Editor

PLOS ONE

 

Additional Editor Comments (optional):

Dear Dr. Bowman,

Thank you for addressing the comments raised by the three reviewers. I believe the manuscript has been sufficiently improved based on their suggestions.

I do not have any further comments.

Best,

Praveen

 

Reviewers' comments: