Myosin Va binding to neurofilaments is essential for correct myosin Va distribution and transport and neurofilament density.
The identification of molecular motors that modulate the neuronal cytoskeleton has been elusive. Here, we show that a molecular motor protein, myosin Va, is present in high proportions in the cytoskeleton of mouse CNS and peripheral nerves. Immunoelectron microscopy, coimmunoprecipitation, and blot overlay analyses demonstrate that myosin Va in axons associates with neurofilaments, and that the NF-L subunit is its major ligand. A physiological association is indicated by observations that the level of myosin Va is reduced in axons of NF-L-null mice lacking neurofilaments and increased in mice overexpressing NF-L, but unchanged in NF-H-null mice. In vivo pulse-labeled myosin Va advances along axons at slow transport rates overlapping with those of neurofilament proteins and actin, both of which coimmunoprecipitate with myosin Va. Eliminating neurofilaments from mice selectively accelerates myosin Va translocation and redistributes myosin Va to the actin-rich subaxolemma and membranous organelles. Finally, peripheral axons of dilute-lethal mice, lacking functional myosin Va, display selectively increased neurofilament number and levels of neurofilament proteins without altering axon caliber. These results identify myosin Va as a neurofilament-associated protein, and show that this association is essential to establish the normal distribution, axonal transport, and content of myosin Va, and the proper numbers of neurofilaments in axons.
Rao
Mala V
MV
Center for Dementia Research, Nathan Kline Institute, NYU School of Medicine, 140 Old Orangeburg Road, Orangeburg, NY 10962, USA.
Engle
Linda J
LJ
Mohan
Panaiyur S
PS
Yuan
Aidong
A
Qiu
Dike
D
Cataldo
Anne
A
Hassinger
Linda
L
Jacobsen
Stephen
S
Lee
Virginia M-Y
VM
Andreadis
Athena
A
Julien
Jean-Pierre
JP
Bridgman
Paul C
PC
Nixon
Ralph A
RA
D000818:Animals; D001370:Axonal Transport / Q000502:physiology*; D001369:Axons / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D001419:Bacteria; D003599:Cytoskeleton / Q000378:metabolism; D007382:Intermediate Filaments / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016253:Microscopy, Immunoelectron; D020409:Molecular Motor Proteins / Q000378:metabolism; D024701:Myosin Type V / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012584:Sciatic Nerve / Q000378:metabolism
D020409:Molecular Motor Proteins; D016900:Neurofilament Proteins; D011993:Recombinant Fusion Proteins; C044882:neurofilament protein L; D024701:Myosin Type V