Chemical-genetic inhibition of a sensitized mutant myosin Vb demonstrates a role in peripheral-pericentriolar membrane traffic.
Selective, in situ inhibition of individual unconventional myosins is a powerful approach to determine their specific physiological functions. Here, we report the engineering of a myosin Vb mutant that still hydrolyzes ATP, yet is selectively sensitized to an N(6)-substituted ADP analog that inhibits its activity, causing it to remain tightly bound to actin. Inhibition of the sensitized mutant causes inhibition of accumulation of transferrin in the cytoplasm and increases levels of plasma-membrane transferrin receptor, suggesting that myosin Vb functions in traffic between peripheral and pericentrosomal compartments.
Provance
D William
DW
McLaughlin Research Institute, Great Falls, MT 59405, USA.
Gourley
Christopher R
CR
Silan
Colleen M
CM
Cameron
L C
LC
Shokat
Kevan M
KM
Goldenring
James R
JR
Shah
Kavita
K
Gillespie
Peter G
PG
Mercer
John A
JA
D000199:Actins / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002502:Centrioles / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D024701:Myosin Type V / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering; D021381:Protein Transport; D051381:Rats; D011990:Receptors, Transferrin / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism
D000199:Actins; D011990:Receptors, Transferrin; D014168:Transferrin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D024701:Myosin Type V