Structural modules of the large subunits of RNA polymerase. Introducing archaebacterial and chloroplast split sites in the β and β' subunits of Escherichia coli RNA polymerase

Konstantin Severinov, Arkady Mustaev, Aleksandr Kukarin, Oriana Muzzin, Irina Bass, Seth A. Darst, Alex Goldfarb

Research output: Contribution to journalArticlepeer-review

72 Citations (Scopus)

Abstract

The β and β' subunits of Escherichia coli DNA-dependent RNA polymerase are highly conserved throughout eubacterial and eukaryotic kingdoms. However, in some archaebacteria and chloroplasts, the corresponding sequences are 'split' into smaller polypeptides that are encoded by separate genes. To test if such split sites can be accommodated into E. coli RNA polymerase, subunit fragments encoded by the segments of E. coli rpoB and rpoC genes corresponding to archaebacterial and chloroplast split subunits were individually overexpressed. The purified fragments, when mixed in vitro with complementing intact RNA polymerase subunits, yielded an active enzyme capable of catalyzing the phosphodiester bond formation. Thus, the large subunits of eubacteria and eukaryotes are composed of independent structural modules corresponding to the smaller subunits of archaebacteria and chloroplasts.

Original languageEnglish
Pages (from-to)27969-27974
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number44
DOIs
Publication statusPublished - 1996
Externally publishedYes

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