PONE-D-24-18608Fasciclin 2 functions as an expression-level switch on EGFR to control organ shape and size in DrosophilaPLOS ONE

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 [This work was supported by PID2021-123407OB-I00, CEX2021-001165-S and BFU2016-76295-R from MCIN/AEI/10.13039/501100011033 and by “ERDF a way of making Europe”; PROMETEU 2021-027 from Generalitat Valenciana, and SAF2004-06593 from MCYT.].  

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: In this manuscript, the author has studied how Fasciclin 2, a cell adhesion molecule, controls organ size in fruit flies using wing disc as a model system. The author has used sophisticated genetic tools and techniques along with imaging studies to show a modulatory role of this gene on EGFR signaling. While the gain-of-function assays indicates a role of Fasciclin 2 as a non-cell autonomous repressor of EGFR, the loss-of-function assays indicate an interaction of this gene with other IgCAMs such as Fipi and Elff. It is interesting that depending on the specific cellular context, Fasciclin 2 can act cell autonomously as well as non-cell autonomously which may impact cell proliferation rate and organ size. Overall, this study is interesting and well done. Here are my suggestions:

1. The author may want to reorganize Fig1 and Fig.2 for better representation. Fig. 1A, B, D, F, I, and Fig. 2 A, C, F may be accommodated in Fig. 1. Rest should go to Fig. 2.

2. In Fig. 7C-D, the label written in green is not clearly legible. The author should use yellow or some other color so that the label is legible.

3. Also, do the author know the efficiency of the RNAi lines? Are all these lines published? If not, the author may want to test the efficiency of those lines.

4. Scale bar size should be written in the Figure legend for Fig. 1C.

5. Scale bar missing in Fig. 1G, 7B-D, S1B-D, and Fig. S2.

Reviewer #2: In this study, Fasciclin 2 (Fas2) emerges as a pivotal protein in the development of imaginal discs in Drosophila, which are precursors to adult structures like wings and the head capsule. Fas2 is shown to play a crucial role in normal development, but its overexpression causes significant effects on organ size and shape which is known from previous studies.

Overexpression of Fas2 results in a dose-dependent reduction in the size of adult organs, particularly wings and the head capsule. This overexpression also caused abnormal shapes in these organs, emphasizing the need for precise regulation of Fas2 levels during development.

Interestingly, the effects of Fas2 overexpression are non-cell autonomous, influencing neighbouring normal tissues as well. It also reduced their size without inducing cell death or activating common growth-regulating pathways like the JNK signaling pathway.

The data points out Fas2 overexpression represses the activity of EGFR (Epidermal Growth Factor Receptor) in neighbouring cells. This repression of EGFR is crucial for the observed reduction in organ size, indicating that Fas2's regulatory role involves interactions with EGFR signaling pathways.

Fipi and Elff, interact with Fas2 and are essential for its repressive function on EGFR at high expression levels. These interactions highlight the complexity of Fas2's regulatory network, where specific molecules like IgCAMs mediate its effects on EGFR signaling.

This manuscript provides insights into the multifaceted role of Fas2 in Drosophila development, particularly in regulating organ size and shape through its interactions with EGFR signaling pathways. The findings suggest a sophisticated regulatory network involving both cell-autonomous and non-cell autonomous mechanisms, mediated by specific protein interactions. Understanding these mechanisms not only sheds light on fundamental developmental processes but also underscores the importance of precise molecular regulation in ensuring proper tissue morphogenesis. It will be interesting to see how Fas2, and other CAMs engage in alternate homo or heteromeric interactions to regulate size and shape. Commend the authors on this decent study.

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Reviewer #1: No

Reviewer #2: No

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Fasciclin 2 functions as an expression-level switch on EGFR to control organ shape and size in Drosophila

PONE-D-24-18608R1

Dear Dr. Garcia-Alonso,

We’re pleased to inform you that your manuscript has been reviewed scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

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Kind regards,

Ashutosh Pandey, Ph.D.

Academic Editor

PLOS ONE

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