RNA polymerase from Bacillus subtilis: isolation of core and holo enzyme by DNA-cellulose chromatography.

Abstract:

:A new procedure for the purification of B. subtilis RNA polymerase, based on mild lysis of cells, low speed centrifugation, gel filtration, DEAE-Sephadex chromatography and affinity chromatography on DNA-cellulose, yields three forms of enzyme referred here as enzyme A, B and C. As revealed by SDS gel electrophoresis, enzyme A has the subunit structure of core polymerase plus some small polypeptides. Its catalytic properties are similar to those of core polymerase. Enzyme B has the composition of core polymerase. Both enzymes A and B can be stimulated by the addition of beta factor. Enzyme C has the holo-enzyme composition. The pattern of sensitivity of the three forms of enzyme towards KCl are very different: enzymes A and B, even at low concentration of salt, are inhibited with all the DNA templates tested, whereas enzyme C shows a pattern of stimulation specific for each DNA tested. The transcripts of the three enzymes on phage SPP1 DNA template have been analyzed by hybridization to the separated strands. Only enzyme C selectively transcribed the H strands.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Plevan P,Albertini AM,Galizzi A,Adamoli A,Mastromei G,Riva S,Cassani G

doi

10.1093/nar/4.3.603

subject

Has Abstract

pub_date

1977-03-01 00:00:00

pages

603-23

issue

3

eissn

0305-1048

issn

1362-4962

journal_volume

4

pub_type

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