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Published January 2015 | Accepted Version + Supplemental Material
Journal Article Open

Regulation of the formin cappuccino is critical for polarity of Drosophilao ocytes

Abstract

The Drosophila formin Cappuccino (Capu) creates an actin mesh‐like structure that traverses the oocyte during midoogenesis. This mesh is thought to prevent premature onset of fast cytoplasmic streaming which normally happens during late‐oogenesis. Proper cytoskeletal organization and cytoplasmic streaming are crucial for localization of polarity determinants such as osk, grk, bcd, and nanos mRNAs. Capu mutants disrupt these events, leading to female sterility. Capu is regulated by another nucleator, Spire, as well as by autoinhibition in vitro. Studies in vivo confirm that Spire modulates Capu's function in oocytes; however, how autoinhibition contributes is still unclear. To study the role of autoinhibition in flies, we expressed a Capu construct that is missing the Capu Inhibitory Domain, CapuΔN. Consistent with a gain of activity due to loss of autoinhibition, the actin mesh was denser in CapuΔN oocytes. Further, cytoplasmic streaming was delayed and fertility levels decreased. Localization of osk mRNA in early stages, and bcd and nanos in late stages, were disrupted in CapuΔN‐expressing oocytes. Finally, evidence that these phenotypes were due to a loss of autoinhibition comes from coexpression of the N‐terminal half of Capu with CapuΔN, which suppressed the defects in actin, cytoplasmic streaming and fertility. From these results, we conclude that Capu can be autoinhibited during Drosophila oocyte development.

Additional Information

© 2014 Wiley Periodicals, Inc. We thank the Gavis, Schüpbach and St. Johnston labs for generously sharing fly lines used in our experiments. We thank the De Robertis lab for use of their dark field microscope and the Courey lab for reagents and help with methods. B.B. thanks his parents (Bor Batsuuri and Altanchimeg Mishig) for bringing him to this world and to this country. This work was supported by several funding sources: UCLA Whitcome fellowship (B.B.); National Institutes of Health (R01GM096133), Burroughs-Welcome Fund (Career Award in the Biomedical Sciences), and March of Dimes Foundation (#1-FY12-442) (M.E.Q.). Stocks obtained from the Bloomington Drosophila Stock Center (NIH P4OD018537) were used in this study. The authors declare no competing financial interests.

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Accepted Version - nihms-652015.pdf

Supplemental Material - cm21205-sup-0001-suppinfo.docx

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August 20, 2023
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