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)


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
Issue number10
Publication statusPublished - 9 May 1995
Externally publishedYes


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