ADPribosylation of nuclear proteins labeled with [3H]adenosine: Changes during the HeLa cell cycle

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Abstract

Cell cycle variations in the modification of histones and nonhistones by ADPribosylation were investigated. Proteins of HeLa interphase nuclei and metaphase chromosomes were radioactively labeled in vivo with [3H]adenosine. Histones of metaphase chromosomes were extensively modified by ADPribosylation, with H2B, H2A and H4 being predominant acceptors of [3H]adenosine label. For histones of interphase nuclei from synchronized cells, the highest level of 3H labeling was observed by two-dimensional gel electrophoresis to occur in S phase. The minimum level was noted in G1 phase. ADPribosylation of histones is, however, significant during all phases of the cell cycle. These conclusions were confirmed by experiments using [32P]NAD. The results with the specific inhibitor of ADPribosylation, 3-aminobenzamide, and with snake venom phosphodiesterase indicated that the radioactive isotopes were incorporated as ADPribose. Two-dimensional gels of HeLa nonhistones labeled with [3H]adenosine showed strikingly different patterns for interphase and metaphase samples. Over 100 ADPribosylated species were found for interphase nuclei, but poly(ADPribose) polymerase was the only major acceptor for metaphase chromosomes. A simple pattern was also revealed for nuclear scaffolds, with the 'lamins' and poly(ADPribose) polymerase being identifiable as modified species.

Original languageEnglish (US)
Pages (from-to)222-230
Number of pages9
JournalBBA - Gene Structure and Expression
Volume909
Issue number3
DOIs
StatePublished - Aug 25 1987

Bibliographical note

Funding Information:
This researchw as supportedb y grants from the Minnesota Medical Foundation and the National Instituteso f Health.

Keywords

  • ADPribosylation
  • Adenosine
  • HeLa cell cycle
  • Histone
  • Nonhistone
  • [H]Adenosine label

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