TY - JOUR
T1 - Comprehensive knockout analysis of the Rab family GTPases in epithelial cells
AU - Homma, Yuta
AU - Kinoshita, Riko
AU - Kuchitsu, Yoshihiko
AU - Wawro, Paulina S.
AU - Marubashi, Soujiro
AU - Oguchi, Mai E.
AU - Ishida, Morié
AU - Fujita, Naonobu
AU - Fukuda, Mitsunori
N1 - Funding Information:
This work was supported in part by the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant-in-Aid for Young Scientists, 18K14692 to Y. Homma; Grant-in-Aid for Scientific Research (C), 18K06202 to N. Fujita; Grant-in-Aid for Scientific Research (B), 15H04367 to M. Fukuda; Grant-in-Aid for Scientific Research on Innovative Areas, 17H05682 to M. Fukuda), by the Japan Science and Technology Agency Core Research for Evolutional Science and Technology (grant JPMJCR17H4 to M. Fukuda), by the Japan Society for the Promotion of Science (M.E. Oguchi and S. Marubashi), and by the Division for Interdisciplinary Advanced Research and Education, Tohoku University (to Y. Kuchitsu and S. Marubashi). The authors declare no competing financial interests.
PY - 2019/6/3
Y1 - 2019/6/3
N2 - 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.
AB - 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.
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U2 - 10.1083/JCB.201810134
DO - 10.1083/JCB.201810134
M3 - Article
C2 - 31072826
AN - SCOPUS:85067214269
VL - 218
SP - 2035
EP - 2050
JO - Journal of Cell Biology
JF - Journal of Cell Biology
SN - 0021-9525
IS - 6
ER -