ADP-ribosylation of nonhistone high mobility group proteins in intact cells.
The ADP-ribosylation of nonhistone, high mobility group (HMG) proteins in intact cultured cells was investigated. Radioactively labeled adenosine was used as a precursor to detect (ADP-ribose)n on protein. A protein fraction enriched in HMG proteins and histone H1 was separated from RNA and DNA by CsCl density gradient centrifugation, 5% perchloric acid extraction, and CM-Sephadex C-50 column chromatography. Poly- and mono(ADP-ribose) were recovered following alkaline hydrolysis, and 5'-AMP and (2'-(5"-phosphoribosyl)-5'-AMP) were produced by phosphodiesterase treatment, indicating that the protein-bound radioactive material was (ADP-ribose)n. An average chain length of 1.5 to 1.8 was determined. Analysis of proteins by sodium dodecyl sulfate and acetic acid/urea polyacrylamide gel electrophoresis demonstrated that HMG 1, 2, 14, and 17 as well as histone H1 contained (ADP-ribose)n. Treatment of cells with 3-aminobenzamide, an inhibitor of (ADP-ribose)n synthetase, decreased endogenous ADP-ribosylation in both types of chromosomal proteins but that of HMG 14 and 17 was affected more.
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D005260:Female; D006609:High Mobility Group Proteins; D008325:Mammary Neoplasms, Experimental; D051379:Mice; D008809:Mice, Inbred C3H; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism
D002868:Chromosomal Proteins, Non-Histone; D006609:High Mobility Group Proteins; D009702:Nucleoside Diphosphate Sugars; D000246:Adenosine Diphosphate Ribose; D011064:Poly Adenosine Diphosphate Ribose