A Genome-wide RNAi Screen for Microtubule Bundle Formation and Lysosome Motility Regulation in Drosophila S2 Cells

Amber L. Jolly, Chi Hao Luan, Brendon E. Dusel, Sara F. Dunne, Michael Winding, Vishrut J. Dixit, Chloe Robins, Jennifer L. Saluk, David J. Logan, Anne E. Carpenter, Manu Sharma, Deborah Dean, Andrew R. Cohen*, Vladimir I. Gelfand

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Long-distance intracellular transport of organelles, mRNA, and proteins ("cargo") occurs along the microtubule cytoskeleton by the action of kinesin and dynein motor proteins, but the vast network of factors involved in regulating intracellular cargo transport are still unknown. We capitalize on the Drosophila melanogaster S2 model cell system to monitor lysosome transport along microtubule bundles, which require enzymatically active kinesin-1 motor protein for their formation. We use an automated tracking program and a naive Bayesian classifier for the multivariate motility data to analyze 15,683 gene phenotypes and find 98 proteins involved in regulating lysosome motility along microtubules and 48 involved in the formation of microtubule filled processes in S2 cells. We identify innate immunity genes, ion channels, and signaling proteins having a role in lysosome motility regulation and find an unexpected relationship between the dynein motor, Rab7a, and lysosome motility regulation.

Original languageEnglish (US)
Pages (from-to)611-620
Number of pages10
JournalCell reports
Volume14
Issue number3
DOIs
StatePublished - Jan 26 2016

Funding

We would like to thank Pradeep Sivakumar (Northwestern University, Evanston, IL), Mark Bray, and the Carpenter lab (Broad Institute, MA) for help with batch image data processing. The project described was supported in part by the H Foundation Cancer Research Fund and Robert H. Lurie Comprehensive Cancer Center. The content is solely the responsibility of the authors and does not necessarily represent the official views of the H Foundation and Robert H. Lurie Comprehensive Cancer Center. Additional funding was provided by a National Science Foundation Graduate Research Fellowship Program fellowship to A.L.J. and National Institute of General Medical Science/National Institutes of Health grants R01 GM052111 to V.I.G. and R01 GM089652 to A.E.C. We would like to thank Pradeep Sivakumar (Northwestern University, Evanston, IL), Mark Bray, and the Carpenter lab (Broad Institute, MA) for help with batch image data processing. The project described was supported in part by the H Foundation Cancer Research Fund and Robert H. Lurie Comprehensive Cancer Center. The content is solely the responsibility of the authors and does not necessarily represent the official views of the H Foundation and Robert H. Lurie Comprehensive Cancer Center. Additional funding was provided by a National Science Foundation Graduate Research Fellowship Program fellowship to A.L.J. and National Institute of General Medical Science/National Institutes of Health grants R01 GM052111 to V.I.G. and R01 GM089652 to A.E.C.

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology

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