title: A Genome-wide RNAi Screen for Microtubule Bundle Formation and Lysosome Motility Regulation in Drosophila S2 Cells.
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.
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University, Chicago, IL 60611, USA; Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA.
authors.forename: Amber L
authors.initials: AL
authors.lastname: Jolly
authors.affiliation: High Throughput Analysis Laboratory, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Chi-Hao
authors.initials: CH
authors.lastname: Luan
authors.affiliation: High Throughput Analysis Laboratory, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Brendon E
authors.initials: BE
authors.lastname: Dusel
authors.affiliation: High Throughput Analysis Laboratory, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Sara F
authors.initials: SF
authors.lastname: Dunne
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Winding
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University, Chicago, IL 60611, USA.
authors.forename: Vishrut J
authors.initials: VJ
authors.lastname: Dixit
authors.affiliation: High Throughput Analysis Laboratory, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Chloe
authors.initials: C
authors.lastname: Robins
authors.affiliation: High Throughput Analysis Laboratory, Northwestern University, Evanston, IL 60208, USA.
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Saluk
authors.affiliation: Imaging Platform, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
authors.forename: David J
authors.initials: DJ
authors.lastname: Logan
authors.affiliation: Imaging Platform, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA.
authors.forename: Anne E
authors.initials: AE
authors.lastname: Carpenter
authors.affiliation: Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA.
authors.forename: Manu
authors.initials: M
authors.lastname: Sharma
authors.affiliation: Children's Hospital Oakland Research Institute, Oakland, CA 94609, USA.
authors.forename: Deborah
authors.initials: D
authors.lastname: Dean
authors.affiliation: Department of Electrical and Computer Engineering, Drexel University, Philadelphia, PA 19104, USA. Electronic address: acohen@coe.drexel.edu.
authors.forename: Andrew R
authors.initials: AR
authors.lastname: Cohen
authors.affiliation: Department of Cell and Molecular Biology, Northwestern University, Chicago, IL 60611, USA. Electronic address: vgelfand@northwestern.edu.
authors.forename: Vladimir I
authors.initials: VI
authors.lastname: Gelfand
mesh_terms: D000818:Animals; D001499:Bayes Theorem; D002478:Cells, Cultured; D004330:Drosophila / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004398:Dyneins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D016678:Genome; D008247:Lysosomes / Q000502:physiology*; D008870:Microtubules / Q000378:metabolism*; D010641:Phenotype; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D059008:Time-Lapse Imaging; D020691:rab GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
chemical_list: D029721:Drosophila Proteins; D012330:RNA, Double-Stranded; D000090802:rab7 GTP-Binding Proteins; D004398:Dyneins; D020691:rab GTP-Binding Proteins