@article{6bb4ed5c24e2490ea54914e0b3ead4db,
title = "Phosphorylation controls spatial and temporal activities of motor-PRC1 complexes to complete mitosis",
abstract = "During mitosis, spindle architecture alters as chromosomes segregate into daughter cells. The microtubule crosslinker protein regulator of cytokinesis 1 (PRC1) is essential for spindle stability, chromosome segregation and completion of cytokinesis, but how it recruits motors to the central spindle to coordinate the segregation of chromosomes is unknown. Here, we combine structural and cell biology approaches to show that the human CENP-E motor, which is essential for chromosome capture and alignment by microtubules, binds to PRC1 through a conserved hydrophobic motif. This binding mechanism is also used by Kinesin-4 Kif4A:PRC1. Using in vitro reconstitution, we demonstrate that CENP-E slides antiparallel PRC1-crosslinked microtubules. We find that the regulation of CENP-E -PRC1 interaction is spatially and temporally coupled with relocalization to overlapping microtubules in anaphase. Finally, we demonstrate that the PRC1-microtubule motor interaction is essential in anaphase to control chromosome partitioning, retain central spindle integrity and ensure cytokinesis. Taken together our findings reveal the molecular basis for the cell cycle regulation of motor-PRC1 complexes to couple chromosome segregation and cytokinesis.",
keywords = "kinesin, mitosis, spindle, microtubule, phosphorylation",
author = "Agata Gluszek-Kustusz and Benjamin Craske and Thibault Legal and Toni Mchugh and Welburn, {Julie P I}",
note = "Funding Information: His‐GFP‐PRC1 and His‐PRC1 were kind gifts from Tarun Kapoor (Rockfeller University). We are grateful to Iain Cheeseman (Whitehead institute for Biomedical Research, Cambridge, Massachusetts, USA) for sharing the human HeLa‐inducible CRISPR/Cas9/PRC1‐G2.2 knock‐out (KO) cell line. We thank Owen Davies for help with AlphaFold2 and Liz Blackburn for help with ITC. We thank Martin Wear for help with SEC‐MALS, and COIL for imaging support. JPIW received a JEDI award from the Life Science Editors Foundation, which provided editorial feedback on an early draft of the manuscript. JPIW is supported by a Wellcome Senior Research Fellowship (207430). BC was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) (grant number BB/M010996/1). This project was also supported by a COVID‐reboot grant from the Royal Society of Edinburgh. The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust (203149). A Multi‐User Equipment grant for the Edinburgh Protein Production Facility is also funded by the Wellcome Trust (101527). Funding Information: His-GFP-PRC1 and His-PRC1 were kind gifts from Tarun Kapoor (Rockfeller University). We are grateful to Iain Cheeseman (Whitehead institute for Biomedical Research, Cambridge, Massachusetts, USA) for sharing the human HeLa-inducible CRISPR/Cas9/PRC1-G2.2 knock-out (KO) cell line. We thank Owen Davies for help with AlphaFold2 and Liz Blackburn for help with ITC. We thank Martin Wear for help with SEC-MALS, and COIL for imaging support. JPIW received a JEDI award from the Life Science Editors Foundation, which provided editorial feedback on an early draft of the manuscript. JPIW is supported by a Wellcome Senior Research Fellowship (207430). BC was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) (grant number BB/M010996/1). This project was also supported by a COVID-reboot grant from the Royal Society of Edinburgh. The Wellcome Centre for Cell Biology is supported by core funding from the Wellcome Trust (203149). A Multi-User Equipment grant for the Edinburgh Protein Production Facility is also funded by the Wellcome Trust (101527). Publisher Copyright: {\textcopyright} 2023 The Authors. Published under the terms of the CC BY 4.0 license.",
year = "2023",
month = nov,
day = "2",
doi = "10.15252/embj.2023113647",
language = "English",
volume = "42",
journal = "The EMBO journal",
issn = "0261-4189",
publisher = "Wiley-Blackwell",
number = "21",
}