Acetylation of RNA processing proteins and cell cycle proteins in mitosis.
Mitosis is a highly regulated process in which errors can lead to genomic instability, a hallmark of cancer. During this phase of the cell cycle, transcription is silent and RNA translation is inhibited. Thus, mitosis is largely driven by post-translational modification of proteins, including phosphorylation, methylation, ubiquitination, and sumoylation. Here, we show that protein acetylation is prevalent during mitosis. To identify proteins that are acetylated, we synchronized HeLa cells in early prometaphase and immunoprecipitated lysine-acetylated proteins with antiacetyl-lysine antibody. The immunoprecipitated proteins were identified by LC-ESI-MS/MS analysis. These include proteins involved in RNA translation, RNA processing, cell cycle regulation, transcription, chaperone function, DNA damage repair, metabolism, immune response, and cell structure. Immunoprecipitation followed by Western blot analyses confirmed that two RNA processing proteins, eIF4G and RNA helicase A, and several cell cycle proteins, including APC1, anillin, and NudC, were acetylated in mitosis. We further showed that acetylation of APC1 and NudC was enhanced by apicidin treatment, suggesting that their acetylation was regulated by histone deacetylase. Moreover, treating mitotic cells with apicidin or trichostatin A induced spindle abnormalities and cytokinesis failure. These studies suggest that protein acetylation/deacetylation is likely an important regulatory mechanism in mitosis.
Chuang
Carol
C
Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, Texas 77030, USA.
Lin
Sue-Hwa
SH
Huang
Feilei
F
Pan
Jing
J
Josic
Djuro
D
Yu-Lee
Li-yuan
LY
D000107:Acetylation; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D048749:Cytokinesis / Q000187:drug effects; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D015151:Immunoblotting; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008938:Mitosis / Q000187:drug effects / Q000502:physiology*; D010456:Peptides, Cyclic / Q000494:pharmacology; D040901:Proteomics; D012323:RNA Processing, Post-Transcriptional; D013347:Subcellular Fractions / Q000737:chemistry; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism
D018797:Cell Cycle Proteins; D006877:Hydroxamic Acids; D010456:Peptides, Cyclic; D014404:Tubulin; C102351:apicidin; C012589:trichostatin A; D004247:DNA
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