The distribution and turnover of labeled methyl groups in histone fractions of cultured mammalian cells |
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| Authors: | P Byvoet G R Shepherd J M Hardin B J Noland |
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| Affiliation: | 1. Department of Developmental and Molecular Biology, Montefiore Hospital, Bronx, NY;2. Gottesman Institute of Stem Cell Biology and Regenerative Medicine, Montefiore Hospital, Bronx, NY;3. Department of Cell Biology, Montefiore Hospital, Bronx, NY;4. Department of Pediatrics, Montefiore Hospital, Bronx, NY;5. Department of Medicine (Oncology), Albert Einstein College of Medicine, Bronx, NY;1. Department of Surgery and Biomedical Sciences, Cedars-Sinai Medical Center, Los Angeles, CA 90048, United States;2. Department of Surgery, University of Michigan Medical Center, Ann Arbor, MI, 48109, United States;3. Key Laboratory of Genomics and Precision Medicine, China Gastrointestinal Cancer Research Center, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, 100101, China;4. College of Material Chemistry and Chemical Engineering, Hangzhou Normal University, Hangzhou, 310036, China;5. Department of Toxicology, School of Public Health, Shenyang Medical College, Liaoning, 110034, China;6. Tianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 300193, China;7. State Key Laboratory of Protein and Plant Gene Research, College of Life Sciences, Peking University, Beijing, 100871, China;1. Shanghai Institute of Nutrition and Health, Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, 200031, China;2. School of Public Health, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China;3. Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, China;4. School of Public Health, Guangzhou Medical University, Guangdong, 511436, China;1. SCIMS, University of the South Pacific, Laucala Bay, Suva, Fiji;2. CS, University of New Orleans, LA, USA;3. CIS, University of Massachusetts Dartmouth, MA, USA;4. IIIS, Griffith University, Brisbane, QLD, Australia |
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| Abstract: | Random cultures of mammalian cells were labeled with [14C-Me]-methionine. Cells were harvested either directly after labeling or after varying periods of further growth in unlabeled medium. Harvested cells were fractionated into cytosol, nucleoplasm, histone, and residual protein fractions. The distribution of radiomethyl activity among the amino acids of each fraction was determined by hydrolysis and ion-exchange chromatography, while the distribution of volatile radiomethyl components was determined by hydrolysis and gas chromatography. The specific activities of methylated lysine residues were found to be considerably higher in histones than in any other fraction, while the nucleoplasm and residual proteins contained methyl arginine at higher specific activities than did the histones. Among the histone fractions, only F2b was found to contain methylated arginine. Among the fractions, only histones were found to contain methanol as their main volatile component after brief alkaline hydrolysis. A comparison of the half-lives of histone fractions and their methyl lysine and methyl arginine components indicated that histone methylation was irreversible. |
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