title: EB1 acetylation by P300/CBP-associated factor (PCAF) ensures accurate kinetochore-microtubule interactions in mitosis.
abstract: In eukaryotes, microtubules are essential for cellular plasticity and dynamics. Here we show that P300/CBP-associated factor (PCAF), a kinetochore-associated acetyltransferase, acts as a negative modulator of microtubule stability through acetylation of EB1, a protein that controls the plus ends of microtubules. PCAF acetylates EB1 on K220 and disrupts the stability of a hydrophobic cavity on the dimerized EB1 C terminus, which was previously reported to interact with plus-end tracking proteins (TIPs) containing the SxIP motif. As determined with an EB1 acetyl-K220-specific antibody, K220 acetylation is dramatically increased in mitosis and localized to the spindle microtubule plus ends. Surprisingly, persistent acetylation of EB1 delays metaphase alignment, resulting in impaired checkpoint silencing. Consequently, suppression of Mad2 overrides mitotic arrest induced by persistent EB1 acetylation. Thus, our findings identify dynamic acetylation of EB1 as a molecular mechanism to orchestrate accurate kinetochore-microtubule interactions in mitosis. These results establish a previously uncharacterized regulatory mechanism governing localization of microtubule plus-end tracking proteins and thereby the plasticity and dynamics of cells.
authors.affiliation: Anhui Key Laboratory of Cellular Dynamics and Hefei National Laboratory for Physical Sciences at Nanoscale, University of Science and Technology of China School of Life Sciences, Hefei 230027, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Xia
authors.forename: Zhikai
authors.initials: Z
authors.lastname: Wang
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
authors.forename: Bing
authors.initials: B
authors.lastname: Wu
authors.forename: Juncheng
authors.initials: J
authors.lastname: Wang
authors.forename: Tarsha
authors.initials: T
authors.lastname: Ward
authors.forename: Liangyu
authors.initials: L
authors.lastname: Zhang
authors.forename: Xia
authors.initials: X
authors.lastname: Ding
authors.forename: Gary
authors.initials: G
authors.lastname: Gibbons
authors.forename: Yunyu
authors.initials: Y
authors.lastname: Shi
authors.forename: Xuebiao
authors.initials: X
authors.lastname: Yao
mesh_terms: D000107:Acetylation; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D018386:Kinetochores / Q000378:metabolism*; D008239:Lysine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D064246:Mad2 Proteins; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D008938:Mitosis; D008958:Models, Molecular; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity; D050880:p300-CBP Transcription Factors / Q000235:genetics / Q000378:metabolism*; D000099224:p300-CBP-Associated Factor
chemical_list: D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D049452:Green Fluorescent Proteins; D008239:Lysine; D064246:Mad2 Proteins; D008869:Microtubule-Associated Proteins; D012097:Repressor Proteins; D050880:p300-CBP Transcription Factors; D000099224:p300-CBP-Associated Factor; C577697:MAD2L1 protein, human; C500985:MAPRE1 protein, human