Distinct functions of regions 1.1 and 1.2 of RNA polymerase ? subunits from Escherichia coli and Thermus aquaticus in transcription initiation.

ABSTRACT: RNA polymerase (RNAP) from thermophilic Thermus aquaticus is characterized by higher temperature of promoter opening, lower promoter complex stability, and higher promoter escape efficiency than RNAP from mesophilic Escherichia coli. We demonstrate that these differences are in part explained by differences in the structures of the N-terminal regions 1.1 and 1.2 of the E. coli ?(70) and T. aquaticus ?(A) subunits. In particular, region 1.1 and, to a lesser extent, region 1.2 of the E. coli ?(70) subunit determine higher promoter complex stability of E. coli RNAP. On the other hand, nonconserved amino acid substitutions in region 1.2, but not region 1.1, contribute to the differences in promoter opening between E. coli and T. aquaticus RNAPs, likely through affecting the ? subunit contacts with DNA nucleotides downstream of the -10 element. At the same time, substitutions in ? regions 1.1 and 1.2 do not affect promoter escape by E. coli and T. aquaticus RNAPs. Thus, evolutionary substitutions in various regions of the ? subunit modulate different steps of the open promoter complex formation pathway, with regions 1.1 and 1.2 affecting promoter complex stability and region 1.2 involved in DNA melting during initiation.

SUBMITTER: Miropolskaya N 

PROVIDER: S-EPMC3390652 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature