Comprehensive knockout analysis of the Rab family GTPases in epithelial cells

Yuta Homma, Riko Kinoshita, Yoshihiko Kuchitsu, Paulina S. Wawro, Soujiro Marubashi, Mai E. Oguchi, Morié Ishida, Naonobu Fujita, Mitsunori Fukuda

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)2035-2050
Number of pages16
JournalJournal of Cell Biology
Volume218
Issue number6
DOIs
Publication statusPublished - 2019 Jun 3

ASJC Scopus subject areas

  • Cell Biology

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