@article{6e67ec2c4ec34cf0a208da4b99e3d685,
title = "Gatekeeper function for Short stop at the ring canals of the Drosophila ovary",
abstract = "Growth of the Drosophila oocyte requires transport of cytoplasmic materials from the interconnected sister cells (nurse cells) through ring canals, the cytoplasmic bridges that remained open after incomplete germ cell division. Given the open nature of the ring canals, it is unclear how the direction of transport through the ring canal is controlled. In this work, we show that a single Drosophila spectraplakin Short stop (Shot) controls the direction of flow from nurse cells to the oocyte. Knockdown of shot changes the direction of transport through the ring canals from unidirectional (toward the oocyte) to bidirectional. After shot knockdown, the oocyte stops growing, resulting in a characteristic small oocyte phenotype. In agreement with this transport-directing function of Shot, we find that it is localized at the asymmetric actin baskets on the nurse cell side of the ring canals. In wild-type egg chambers, microtubules localized in the ring canals have uniform polarity (minus ends toward the oocyte), while in the absence of Shot, these microtubules have mixed polarity. Together, we propose that Shot functions as a gatekeeper directing transport from nurse cells to the oocyte via the organization of microtubule tracks to facilitate the transport driven by the minus-end-directed microtubule motor cytoplasmic dynein.",
keywords = "Drosophila, actin, cytoskeleton, dynein, intercellular transport, microtubules, molecular motor, oocyte, polarity, spectraplakin",
author = "Wen Lu and Margot Lakonishok and Gelfand, {Vladimir I.}",
note = "Funding Information: We would like to thank Dr. David Glover (Caltech) for ubi-GFP-Pav line, Dr. Derek Applewhite (Reed College) for full-length shot cDNA (shot.LA) and ubi- EB1- GFP line, Dr. Ferenc Jankovics (Institute of Genetics, Biological Research Centre of the Hungarian Academy of Sciences) for shotΔEGC line, Dr. Daniel St. Johnston (University of Cambridge) for mat αtub-RFP-Staufen line, Dr. Vladislav Verkhusha (Albert Einstein College of Medicine) for MoxMaple3 construct, Dr. Michael Welte (University of Rochester) for UASp-GFP-LD line, Dr. Uri Abdu (Ben-Gurion University of the Negev) for UASp-GFP-Patronin line, Dr. Antoine Guichet (CNRS, Institut Jacques Monod) for UASp-EB1-GFP line, the Bloomington Drosophila Stock Center (supported by National Institutes of Health grant P40OD018537) for fly stocks, and Drosophila Genomics Resource Center (supported by National Institutes of Health grant 2P40OD010949) for DNA constructs. The Orb 4H8 monoclonal antibody developed by Dr. Paul D. Schedl's group at Princeton University and anti-Shot mAbRod1 antibody developed by Dr. Peter A. Kolodziej's group at Vanderbilt University were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa. We also thank all the Gelfand laboratory members for support, discussion, and suggestions. W.L. dedicates this work to memory of her college genetics teacher, Professor Zhuo-hua Dai at Peking University. Research reported in this study was supported by the National Institute of General Medical Sciences grants R01GM124029 and R35GM131752 to V.I.G. W.L. M.L. and V.I.G. planned and designed the research. W.L. M.L. and V.I.G. conducted experiments and data analysis; W.L. and V.I.G. wrote the manuscript. The authors declare no competing interests. Funding Information: We would like to thank Dr. David Glover (Caltech) for ubi-GFP-Pav line, Dr. Derek Applewhite (Reed College) for full-length shot cDNA (shot.LA) and ubi- EB1- GFP line, Dr. Ferenc Jankovics (Institute of Genetics, Biological Research Centre of the Hungarian Academy of Sciences) for shot ΔEGC line, Dr. Daniel St. Johnston (University of Cambridge) for mat αtub-RFP-Staufen line, Dr. Vladislav Verkhusha (Albert Einstein College of Medicine) for MoxMaple3 construct, Dr. Michael Welte (University of Rochester) for UASp-GFP-LD line, Dr. Uri Abdu (Ben-Gurion University of the Negev) for UASp-GFP-Patronin line, Dr. Antoine Guichet (CNRS, Institut Jacques Monod) for UASp-EB1-GFP line, the Bloomington Drosophila Stock Center (supported by National Institutes of Health grant P40OD018537 ) for fly stocks, and Drosophila Genomics Resource Center (supported by National Institutes of Health grant 2P40OD010949 ) for DNA constructs. The Orb 4H8 monoclonal antibody developed by Dr. Paul D. Schedl{\textquoteright}s group at Princeton University and anti-Shot mAbRod1 antibody developed by Dr. Peter A. Kolodziej{\textquoteright}s group at Vanderbilt University were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa. We also thank all the Gelfand laboratory members for support, discussion, and suggestions. W.L. dedicates this work to memory of her college genetics teacher, Professor Zhuo-hua Dai at Peking University. Research reported in this study was supported by the National Institute of General Medical Sciences grants R01GM124029 and R35GM131752 to V.I.G. Publisher Copyright: {\textcopyright} 2021 Elsevier Inc.",
year = "2021",
month = aug,
day = "9",
doi = "10.1016/j.cub.2021.05.010",
language = "English (US)",
volume = "31",
pages = "3207--3220.e4",
journal = "Current Biology",
issn = "0960-9822",
publisher = "Cell Press",
number = "15",
}