TY - JOUR
T1 - Role of kinesin-1 - based microtubule sliding in Drosophila nervous system development
AU - Winding, Michael
AU - Kelliher, Michael T.
AU - Lu, Wen
AU - Wildonger, Jill
AU - Gelfand, Vladimir I.
N1 - Funding Information:
We thank Dr. Chris Q. Doe (University of Oregon), Dr. Melissa Rolls (Penn State University), Dr. William M. Saxton (University of California, Santa Cruz), and the Bloomington Stock Center (NIH P40OD018537) for fly stocks. The mouse monoclonal anti-Futsch (22C10) antibody contributed by Seymour Benzer (California Institute of Technology), CSP (DCSP-2, 6D6) monoclonal antibody contributed by Seymour Benzer, and SYT (3H2 2D7) monoclonal antibody contributed by Kai Zinn (California Institute of Technology) were obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at the Department of Biology, University of Iowa. We also thank the Center for Advanced Microscopy at Northwestern University and Constadina Arvanitis for assistance in imaging YFP-Rab5 vesicles in class IV da neurons. The research reported here was supported by the National Institute of General Medical Science of the NIH under Award R01GM052111 (to V.I.G.).
Publisher Copyright:
© 2016, National Academy of Sciences. All rights reserved.
PY - 2016/8/23
Y1 - 2016/8/23
N2 - The plus-end microtubule (MT) motor kinesin-1 is essential for normal development, with key roles in the nervous system. Kinesin-1 drives axonal transport of membrane cargoes to fulfill the metabolic needs of neurons and maintain synapses. We have previously demonstrated that kinesin-1, in addition to its well-established role in organelle transport, can drive MT-MT sliding by transporting "cargo" MTs along "track" MTs, resulting in dramatic cell shape changes. The mechanism and physiological relevance of this MT sliding are unclear. In addition to its motor domain, kinesin-1 contains a second MT-binding site, located at the C terminus of the heavy chain. Here, we mutated this C-terminal MT-binding site such that the ability of kinesin-1 to slide MTs is significantly compromised, whereas cargo transport is unaffected. We introduced this mutation into the genomic locus of kinesin-1 heavy chain (KHC), generating the KhcmutA allele. KhcmutA neurons displayed significant MT sliding defects while maintaining normal transport of many cargoes. Using this mutant, we demonstrated that MT sliding is required for axon and dendrite outgrowth in vivo. Consistent with these results, KhcmutA flies displayed severe locomotion and viability defects. To test the role of MT sliding further, we engineered a chimeric motor that actively slides MTs but cannot transport organelles. Activation of MT sliding in KhcmutA neurons using this chimeric motor rescued axon outgrowth in cultured neurons and in vivo, firmly establishing the role of sliding in axon outgrowth. These results demonstrate that MT sliding by kinesin-1 is an essential biological phenomenon required for neuronal morphogenesis and normal nervous system development.
AB - The plus-end microtubule (MT) motor kinesin-1 is essential for normal development, with key roles in the nervous system. Kinesin-1 drives axonal transport of membrane cargoes to fulfill the metabolic needs of neurons and maintain synapses. We have previously demonstrated that kinesin-1, in addition to its well-established role in organelle transport, can drive MT-MT sliding by transporting "cargo" MTs along "track" MTs, resulting in dramatic cell shape changes. The mechanism and physiological relevance of this MT sliding are unclear. In addition to its motor domain, kinesin-1 contains a second MT-binding site, located at the C terminus of the heavy chain. Here, we mutated this C-terminal MT-binding site such that the ability of kinesin-1 to slide MTs is significantly compromised, whereas cargo transport is unaffected. We introduced this mutation into the genomic locus of kinesin-1 heavy chain (KHC), generating the KhcmutA allele. KhcmutA neurons displayed significant MT sliding defects while maintaining normal transport of many cargoes. Using this mutant, we demonstrated that MT sliding is required for axon and dendrite outgrowth in vivo. Consistent with these results, KhcmutA flies displayed severe locomotion and viability defects. To test the role of MT sliding further, we engineered a chimeric motor that actively slides MTs but cannot transport organelles. Activation of MT sliding in KhcmutA neurons using this chimeric motor rescued axon outgrowth in cultured neurons and in vivo, firmly establishing the role of sliding in axon outgrowth. These results demonstrate that MT sliding by kinesin-1 is an essential biological phenomenon required for neuronal morphogenesis and normal nervous system development.
KW - Axon outgrowth
KW - Dendrite outgrowth
KW - Drosophila
KW - Kinesin-1
KW - Microtubules
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U2 - 10.1073/pnas.1522416113
DO - 10.1073/pnas.1522416113
M3 - Article
C2 - 27512046
AN - SCOPUS:84983610498
VL - 113
SP - E4985-E4994
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 34
ER -