RRC ID 57096
Author Homma Y, Kinoshita R, Kuchitsu Y, Wawro PS, Marubashi S, Oguchi ME, Ishida M, Fujita N, Fukuda M.
Title Comprehensive knockout analysis of the Rab family GTPases in epithelial cells.
Journal J Cell Biol
Abstract The Rab family of small GTPases comprises the largest number of proteins (∼60 in mammals) among the regulators of intracellular membrane trafficking, but the precise function of many Rabs and the functional redundancy and diversity of Rabs remain largely unknown. Here, we generated a comprehensive collection of knockout (KO) MDCK cells for the entire Rab family. We knocked out closely related paralogs simultaneously (Rab subfamily knockout) to circumvent functional compensation and found that Rab1A/B and Rab5A/B/C are critical for cell survival and/or growth. In addition, we demonstrated that Rab6-KO cells lack the basement membrane, likely because of the inability to secrete extracellular matrix components. Further analysis revealed the general requirement of Rab6 for secretion of soluble cargos. Transport of transmembrane cargos to the plasma membrane was also significantly delayed in Rab6-KO cells, but the phenotype was relatively mild. Our Rab-KO collection, which shares the same background, would be a valuable resource for analyzing a variety of membrane trafficking events.
Volume 218(6)
Pages 2035-2050
Published 2019-6-3
DOI 10.1083/jcb.201810134
PII jcb.201810134
PMID 31072826
PMC PMC6548125
MeSH Animals Cell Membrane / metabolism* Dogs Epithelial Cells / cytology* Epithelial Cells / metabolism Gene Knockout Techniques / methods Guanosine Triphosphate / metabolism* HEK293 Cells Humans Intracellular Membranes Madin Darby Canine Kidney Cells Multiprotein Complexes / metabolism* Organelles / physiology* Phenotype Protein Transport RNA, Small Interfering / genetics* rab GTP-Binding Proteins / antagonists & inhibitors* rab GTP-Binding Proteins / genetics rab GTP-Binding Proteins / metabolism
IF 8.891
Times Cited 10
DNA material pMRX-bsr-Myc-Rab2A (RDB17357) pMRX-bsr-Myc-Rab5A (RDB17359) pMRX-puro-EGFP-Rab6A (RDB17361) pMRX-puro-EGFP-Rab11A (RDB17362) pMRX-bsr-Myc-Rab2B (RDB17358) pMRX-puro-EGFP-Rab1B (RDB17360) pMRX-bsr-ss-EGFP-FM4 (RDB17364) pMRX-bsr-ss-EGFP-FM4-TMD (RDB17365) pMRX-bsr-ss-EGFP (RDB17363)