Reconstitution studies show that rifampicin resistance is determined by the largest polypeptide of Bacillus subtilis RNA polymerase

S. M. Halling, K. C. Burtis, R. H. Doi

Research output: Contribution to journalArticle

36 Scopus citations

Abstract

A procedure has been developed to separate the subunits of B. subtilis RNA polymerase rapidly and in good yield. The method involved the use of a blue dextran Sepharose column which bounded the β' subunit. A phosphocellulose column was used to separate the α and β subunits. During purification, the enzyme eluted from the DNA cellulose column in three separate forms in the order α2ββ'δω1, α2ββ'ω1, and α2ββ'ω1σ. Subunit reconstitution studies with RNA polymerase subunits from wild type and a rifampicin resistant mutant indicated that the largest polypeptide was responsible for rifampicin resistance. Thus, this subunit in sodium dodecyl sulfate polyacrylamide gel electrophoresis cannot be used as the sole criterion for designating the functions of the subunits of RNA polymerase.

Original languageEnglish (US)
Pages (from-to)9024-9031
Number of pages8
JournalJournal of Biological Chemistry
Volume252
Issue number24
StatePublished - 1977

ASJC Scopus subject areas

  • Biochemistry

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