Genetic dissection of the transcription cycle. A mutant RNA polymerase that cannot hold onto a promoter.

Eisenacher, Martin; Sagitov, V; Burova, Elena; Nikiforov, Vadim; Goldfarb, Alex

doi:10.1016/s0021-9258(19)88683-0

Cited by 17 publications

(1 citation statement)

References 41 publications

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“…The N-terminal b58/b57 domains include the regions A, B, and C conserved among β homologues from not only prokaryotic but also eukaryotic RNA polymerases (23,24) and the N-terminal-proximal dispensable region (the dispensable region I) between the conserved regions B and C (25). However, no specific function has yet been identified for these N-terminal-proximal regions, except that the region B is one of the four regions forming the binding site of rifampicin (26) and a short deletion in the region C leads to a decrease in the binding activity of RNA polymerase to DNA (27).…”

Section: Resultsmentioning

confidence: 99%

Mapping of Subunit−Subunit Contact Surfaces on the β Subunit of Escherichia coli RNA Polymerase

1999

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The RNA polymerase core enzyme of Escherichia coli is composed of 2alpha, 1beta, and 1beta' subunits. Previously we mapped the alpha-alpha, alpha-beta, and alpha-beta' contact sites on the alpha subunit. Here we analyzed the alpha subunit contact sites on the beta subunit by using various experimental approaches: (i) comparison of the proteolytic cleavage map between the unassembled free beta subunit and the alpha2 beta complex; (ii) analysis of the binary complex formation between His6-tagged intact alpha subunit and various truncated beta fragments; and (iii) analysis of the complex formation between the alpha subunit and various His6-tagged beta fragments. The results altogether indicate that two regions of the beta subunit are involved in the full activity of alpha binding, that is, the primary contact site between residues 737 and 904 and the secondary region with assembly control activity downstream from residue 1138. All of the alpha subunit-beta fragment binary complexes identified in this study were found to bind beta' subunit and form pseudo-core complexes, indicating that the regions of beta involved in alpha subunit contact also participate in interaction with the beta' subunit.

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