Selective and accurate transcription of the Xenopus laevis 5S RNA genes in isolated chromatin by purified RNA polymerase III.
Chromatin isolated from immature oocytes was found to contain an endogenous RNA polymerase activity (RNA nucleotidyltransferase; nucleoside triphosphate:RNA nucleotidyltransferase, EC 2.7.7.6) that synthesizes predominately 5S RNA. However, the levels of total RNA synthesis and 5S RNA synthesis in chromatin were each stimulated 10- to 50-fold by an exogenous RNA polymerase III purified from X. laevis oocytes. The 5S genes in chromatin were transcribed by the exogenous enzyme in a highly selective (3000-fold above random) and predominately asymmetric fashion. A significant fraction of 5S RNA sequences were also found in a discrete transcript, approximately 5S in size. Total RNA synthesis was significantly stimulated when chromatin was transcribed by oocyte RNA polymerase I, murine RNA polymerase II, and low levels of Escherichia coli RNA polymerase. However, these enzymes did not significantly stimulate 5S RNA synthesis above the endogenous levels. Both homologous oocyte RNA polymerase I and III and E. coli RNA polymerase transcribed the 5S genes in deproteinized DNA to approximately the same extent (severalfold above random) and both the sense and anti-sense strands of the gene were transcribed. It appears, therefore, that both chromatin-associated components and a purified RNA polymerase III are necessary and sufficient for the selective and accurate transcription of the 5S RNA genes in vitro.
D000818:Animals; D002474:Cell-Free System; D002843:Chromatin / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D005260:Female; D005796:Genes; D066298:In Vitro Techniques; D007700:Kinetics; D008970:Molecular Weight; D009865:Oocytes; D012320:RNA Polymerase III / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000096:biosynthesis*; D013045:Species Specificity; D013698:Templates, Genetic; D014158:Transcription, Genetic; D014981:Xenopus
D002843:Chromatin; D012335:RNA, Ribosomal; D004247:DNA; D012321:DNA-Directed RNA Polymerases; D012320:RNA Polymerase III
J Mol Biol. 1965 Jul;12(3):829-42
4955314
Cell. 1976 Apr;7(4):477-86
782719
Nature. 1975 Oct 30;257(5529):764-7
1186855
Eur J Biochem. 1975 Aug 15;56(2):335-41
1175627
Biochimie. 1973;55(9):1137-51
4799005
Proc Natl Acad Sci U S A. 1973 Dec;70(12):3440-2
4587253
Proc Natl Acad Sci U S A. 1973 Jul;70(7):2029-32
4516203
J Morphol. 1972 Feb;136(2):153-79
4109871
J Biol Chem. 1974 Jan 10;249(1):249-56
4855627
Biochemistry. 1974 Apr 23;13(9):1896-9
4857734
J Mol Biol. 1973 Oct 25;80(2):229-41
4587403
Proc Natl Acad Sci U S A. 1974 May;71(5):1790-4
4525293
Nat New Biol. 1973 Jan 3;241(105):7-12
4512331
J Biol Chem. 1975 Jan 25;250(2):418-25
1089642
Proc Natl Acad Sci U S A. 1975 Dec;72(12):4835-9
1108006
Science. 1976 Sep 3;193(4256):848-56
948749
Biochemistry. 1976 Apr 6;15(7):1500-9
1259952
J Biol Chem. 1976 Apr 10;251(7):1960-8
773931
J Biol Chem. 1976 Feb 25;251(4):1064-73
1249067
Cell. 1976 Apr;7(4):557-66
954087
Proc Natl Acad Sci U S A. 1975 Jan;72(1):348-52
1054509
Proc Natl Acad Sci U S A. 1974 Nov;71(11):4556-60
4612523
J Mol Biol. 1972 Jun 28;67(3):433-41
4558099
J Mol Biol. 1974 Nov 15;89(4):703-18
4449128
Dev Biol. 1974 Aug;39(2):191-7
4854181
J Mol Biol. 1973 Aug 15;78(3):397-415
4754843