Abstract
Escherichia coli RNA polymerase is composed of four different subunits,α (present in two copies),β, β′ andσ. Among these, theβ′ polypeptide shares nine conserved regions with the largest subunits of eukaryotic RNA polymerases, but its role is poorly understood. We isolated novel mutations in a plasmidborne copy ofrpoC, which encodesβ′, as dominant suppressors of two temperature-sensitivenusA alleles. All 20 suppressors ofnusA11 (single missense mutation) isolated had either of two specific substitutions: Lys for Glu-402 (rpoC10) and Thr for Ala-904 (rpoC111) in theβ′ subunit. In vivo and in vitro transcription assays revealed that therpoC10 allele ofβ′ participates in Rho-dependent transcription termination. On the other hand, of 20 suppressors ofnusA134 (deletion of C-terminal one-third) scattered at 18 distinct sites, 16 were assigned to one of six conserved regions C-I. These results suggested that the conserved domains of theβ′ subunit ofE. coli RNA polymerase are involved in transcript termination or interaction with termination factor(s).
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Communicated by M. Sekiguchi
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Ito, K., Nakamura, Y. Localization ofnusA-suppressing amino acid substitutions in the conserved regions of theβ′ subunit ofEscherichia coli RNA polymerase. Molec. Gen. Genet. 251, 699–706 (1996). https://doi.org/10.1007/BF02174119
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DOI: https://doi.org/10.1007/BF02174119