Myo1c facilitates G-actin transport to the leading edge of migrating endothelial cells.
Addition of actin monomer (G-actin) to growing actin filaments (F-actin) at the leading edge generates force for cell locomotion. The polymerization reaction and its regulation have been studied in depth. However, the mechanism responsible for transport of G-actin substrate to the cell front is largely unknown; random diffusion, facilitated transport via myosin II contraction, local synthesis as a result of messenger ribonucleic acid localization, or F-actin turnover all might contribute. By tracking a photoactivatable, nonpolymerizable actin mutant, we show vectorial transport of G-actin in live migrating endothelial cells (ECs). Mass spectrometric analysis identified Myo1c, an unconventional F-actin-binding motor protein, as a major G-actin-interacting protein. The cargo-binding tail domain of Myo1c interacted with G-actin, and the motor domain was required for the transport. Local microinjection of Myo1c promoted G-actin accumulation and plasma membrane ruffling, and Myo1c knockdown confirmed its contribution to G-actin delivery to the leading edge and for cell motility. In addition, there is no obvious requirement for myosin II contractile-based transport of G-actin in ECs. Thus, Myo1c-facilitated G-actin transport might be a critical node for control of cell polarity and motility.
Department of Cell Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
Yi
Y
Fan
Sandeepa M
SM
Eswarappa
Masahiro
M
Hitomi
Paul L
PL
Fox
D000199:Actins / Q000737:chemistry / Q000502:physiology*; D000818:Animals; D001692:Biological Transport; D002417:Cattle; D002465:Cell Movement / Q000502:physiology*; D022081:Cell Surface Extensions / Q000502:physiology; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology*; D055785:Gene Knockdown Techniques; D024461:Myosin Type I / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D054730:Protein Interaction Domains and Motifs / Q000502:physiology
D000199:Actins; D024461:Myosin Type I