Specific gene transcription in yeast nuclei and chromatin by added homologous RNA polymerases I and II.

P. A. Tekamp, P. Valenzuela, T. Maynard, G. I. Bell, W. J. Rutter

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

Abstract

When treated at pH less than 4.5, yeast nuclei or chromatin lose endogenous RNA synthetic activity. This activity is regained by addition of exogenous RNA polymerases. The specificity of transcription in this system by homologous RNA polymerases I and III has been investigated by gel electrophoresis, hybridization analysis, and RNase T1 mapping. Exogenous RNA polymerase I selectively transcribes rRNA genes. The transcription of these genes by polymerase I is 30- and 8-fold more selective than RNA polymerase III and Escherichia coli polymerase holoenzyme, respectively. Exogenous RNA polymerase III synthesized RNAs similar in size to authentic 5 S RNA, 4.5 S pre-tRNA, and 4 S tRNA. Eleven per cent of this RNA is 5 S RNA as determined by hybridization. Neither polymerase I nor E. coli polymerase synthesizes detectable quantities of RNA in this size range. AT1 ribonuclease digestion of 5 S RNA synthesized by exogenous RNA polymerase III acting on acid-treated chromatin gives a fragment pattern corresponding to that of 5 S RNA. Thus, RNA polymerase III transcribes the entire 5 S gene in this system.

Original languageEnglish
Pages (from-to)955-963
Number of pages9
JournalJournal of Biological Chemistry
Volume254
Issue number3
Publication statusPublished - 10 Feb 1979

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

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