Myosin Vc is a molecular motor that functions in secretory granule trafficking.
Class V myosins are actin-based motor proteins that have critical functions in organelle trafficking. Of the three class V myosins expressed in mammals, relatively little is known about Myo5c except that it is abundant in exocrine tissues. Here we use MCF-7 cells to identify the organelles that Myo5c associates with, image the dynamics of Myo5c in living cells, and test the functions of Myo5c. Endogenous Myo5c localizes to two distinct compartments: small puncta and slender tubules. Myo5c often exhibits a highly polarized distribution toward the leading edge in migrating cells and is clearly distinct from the Myo5a or Myo5b compartments. Imaging with GFP-Myo5c reveals that Myo5c puncta move slowly (approximately 30 nm/s) and microtubule independently, whereas tubules move rapidly (approximately 440 nm/s) and microtubule dependently. Myo5c puncta colocalize with secretory granule markers such as chromogranin A and Rab27b, whereas Myo5c tubules are labeled by Rab8a. TIRF imaging indicates that the granules can be triggered to undergo secretion. To test if Myo5c functions in granule trafficking, we used the Myo5c tail as a dominant negative and found that it dramatically perturbs the distribution of granule markers. These results provide the first live-cell imaging of Myo5c and indicate that Myo5c functions in secretory granule trafficking.
Jacobs
Damon T
DT
Department of Cell and Molecular Physiology, School of Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.
Weigert
Roberto
R
Grode
Kyle D
KD
Donaldson
Julie G
JG
Cheney
Richard E
RE
D000818:Animals; D002460:Cell Line; D053379:Chromogranin A / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D020409:Molecular Motor Proteins / Q000235:genetics / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism; D024701:Myosin Type V / Q000235:genetics / Q000378:metabolism*; D009218:Myosins / Q000235:genetics / Q000378:metabolism; D009478:Neuropeptide Y / Q000235:genetics / Q000378:metabolism; D015739:Nocodazole / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D022142:Secretory Vesicles / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D050257:Tubulin Modulators / Q000494:pharmacology
D053379:Chromogranin A; C459104:MYO5C protein, human; D020409:Molecular Motor Proteins; C101826:Myo5b protein, rat; D009478:Neuropeptide Y; D011993:Recombinant Fusion Proteins; D050257:Tubulin Modulators; C077511:MYO5A protein, human; D024701:Myosin Type V; D018995:Myosin Heavy Chains; D009218:Myosins; D015739:Nocodazole