Assembly of functional Escherichia coli RNA polymerase containing β subunit fragments

Konstantin Severinov, Arkady Mustaev, Elena Severinova, Irina Bass, Mikhail Kashlev, Robert Landick, Vadim Nikiforov, Alex Goldfarb, Seth A. Darst

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

The Escherichia coli rpoB gene, which codes for the 1342-residue β subunit of RNA polymerase (RNAP), contains two dispensable regions centered around codons 300 and 1000. To test whether these regions demarcate domains of the RNAP β subunit, fragments encoded by segments of rpoB flanking the dispensable regions were individually overexpressed and purified. We show that these β-subunit polypeptide fragments, when added with purified recombinant β', σ, and α subunits of RNAP, reconstitute a functional enzyme in vitro. These results demonstrate that the β subunit is composed of at least three distinct domains and open another avenue for in vitro studies of RNAP assembly and structure.

Original languageEnglish
Pages (from-to)4591-4595
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume92
Issue number10
DOIs
Publication statusPublished - 9 May 1995
Externally publishedYes

Fingerprint

Dive into the research topics of 'Assembly of functional Escherichia coli RNA polymerase containing β subunit fragments'. Together they form a unique fingerprint.

Cite this