TY - JOUR
T1 - Rab6a releases LIS1 from a dynein idling complex and activates dynein for retrograde movement
AU - Yamada, Masami
AU - Kumamoto, Kanako
AU - Mikuni, Shintaro
AU - Arai, Yoshiyuki
AU - Kinjo, Masataka
AU - Nagai, Takeharu
AU - Tsukasaki, Yoshikazu
AU - Watanabe, Tomonobu M.
AU - Fukui, Mitsuru
AU - Jin, Mingyue
AU - Toba, Shiori
AU - Hirotsune, Shinji
N1 - Funding Information:
We thank Mika Egami, Yukimi Kira, Mitsuhiro Iwaki, Takamitsu J. Morikawa and Yoriko Yabunaka for their technical support, Hiromichi Nishimura and Keiko Fujimoto for mouse breeding, and Dr Anthony Wynshaw-Boris for his comments on the manuscript. This work was supported by Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan from the Ministry of Education, Science, Sports and Culture of Japan, and Knowledge Cluster Initiative (Stage-2) Research Foundation, to S.H. This work was also supported by Takeda Science Foundation and NOVARTIS Foundation (Japan) for the Promotion of Science to S.H. This work was supported by Grant-in-Aid for Scientific Research (B) of Japan Society for the Promotion of Science (JSPS) (no. 22390056 to M.Y.), Adaptable and Seamless Technology Transfer Program (A-STEP) through Target-driven R&D, Japan Science and Technology Agency (no. AS231Z02224G to M.Y.), Grant-in-Aid for Scientific Research on Innovative Areas of The Ministry of Education, Culture, Sports, Science and Technology (MEXT) (no. 23113723 to M.Y.), the Uehara Memorial Foundation (to M.Y.) and the Cooperative Research Program of ‘Network Joint Research Center for Materials and Devices’ (to M.Y.).
PY - 2013
Y1 - 2013
N2 - Cytoplasmic dynein drives the movement of a wide range of cargoes towards the minus ends of microtubules. We previously demonstrated that LIS1 forms an idling complex with dynein, which is transported to the plus ends of microtubules by kinesin motors. Here we report that the small GTPase Rab6a is essential for activation of idling dynein. Immunoprecipitation and microtubule pull-down assays reveal that the GTP bound mutant, Rab6a(Q72L), dissociates LIS1 from a LIS1-dynein complex, activating dynein movement in in vitro microtubule gliding assays. We monitor transient interaction between Rab6a(Q72L) and dynein in vivo using dual-colour fluorescence cross-correlation spectroscopy in dorsal root ganglion (DRG) neurons. Finally, we demonstrate that Rab6a(Q72L) mediates LIS1 release from a LIS1-dynein complex followed by dynein activation through an in vitro single-molecule assay using triple-colour quantum dots. Our findings reveal a surprising function for GTP bound Rab6a as an activator of idling dynein.
AB - Cytoplasmic dynein drives the movement of a wide range of cargoes towards the minus ends of microtubules. We previously demonstrated that LIS1 forms an idling complex with dynein, which is transported to the plus ends of microtubules by kinesin motors. Here we report that the small GTPase Rab6a is essential for activation of idling dynein. Immunoprecipitation and microtubule pull-down assays reveal that the GTP bound mutant, Rab6a(Q72L), dissociates LIS1 from a LIS1-dynein complex, activating dynein movement in in vitro microtubule gliding assays. We monitor transient interaction between Rab6a(Q72L) and dynein in vivo using dual-colour fluorescence cross-correlation spectroscopy in dorsal root ganglion (DRG) neurons. Finally, we demonstrate that Rab6a(Q72L) mediates LIS1 release from a LIS1-dynein complex followed by dynein activation through an in vitro single-molecule assay using triple-colour quantum dots. Our findings reveal a surprising function for GTP bound Rab6a as an activator of idling dynein.
UR - http://www.scopus.com/inward/record.url?scp=84879610490&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84879610490&partnerID=8YFLogxK
U2 - 10.1038/ncomms3033
DO - 10.1038/ncomms3033
M3 - Article
C2 - 23783758
AN - SCOPUS:84879610490
SN - 2041-1723
VL - 4
JO - Nature communications
JF - Nature communications
M1 - 2033
ER -