Role of the kinetochore/cell cycle checkpoint protein ZW10 in interphase cytoplasmic dynein function
Résumé
Zeste white 10 (ZW10) is a mitotic checkpoint protein and the anchor for cytoplasmic dynein at mitotic kinetochores, though it is expressed throughout the cell cycle. We find that ZW10 localizes to pericentriolar membranous structures during interphase and cosediments with Golgi membranes. Dominant-negative ZW10, anti-ZW10 antibody, and ZW10 RNA interference (RNAi) caused Golgi dispersal. ZW10 RNAi also dispersed endosomes and lysosomes. Live imaging of Golgi, endosomal, and lysosomal markers after reduced ZW10 expression showed a specific decrease in the frequency of minus end-directed movements. Golgi membrane-associated dynein was markedly decreased, suggesting a role for ZW10 in dynein cargo binding during interphase. We also find ZW10 enriched at the leading edge of migrating fibroblasts, suggesting that ZW10 serves as a general regulator of dynein function throughout the cell cycle.
Mots clés
N.I.H
Animals
COS Cells
Cell Cycle Proteins/antagonists & inhibitors/*physiology
Cercopithecus aethiops
Chromosomal Proteins
Non-Histone/antagonists & inhibitors/*physiology
Cytoplasm/*physiology
Dynein ATPase/*physiology
Golgi Apparatus/physiology
Hela Cells
Humans
Interphase/*physiology
Kinetochores/physiology
Microinjections
Microtubule-Associated Proteins/antagonists & inhibitors/*physiology
Organelles/physiology
RNA Interference
Research Support
N.I.H.
Extramural
Non-U.S. Gov't