Functional identification of an anti-sigmaE factor from Thermus thermophilus HB8
Keiko Sakamoto1, Yoshihiro Agari, Shigeyuki Yokoyama
1RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.
Abstract:
The TTHB212 gene from extremely thermophilic bacterium Thermus thermophilus HB8 forms an operon with the upstream sigE gene encoding an extracytoplasmic function sigma factor, sigma(E), the sole alternative sigma factor of this strain, on megaplasmid pTT27. The TTHB212 gene encodes a poorly conserved protein, which has been predicted to be a transmembrane one with N-terminal intracellular and C-terminal extracytoplasmic domains. The N-terminal domain of TTHB212 protein (TTHB212N) prevented sigma(E) from binding to RNA polymerase (RNAP) core enzyme in vitro, and TTHB212N bound sigma(E) in a molar ratio of 1:1 when both proteins were co-expressed in Escherichia coli. Furthermore, TTHB212N inhibited the transcription activity of RNAP-sigma(E) holoenzyme, but not that of the RNAP-sigma(A) one, in vitro. The expression of several genes that are under the control of sigma(E) was increased in a TTHB212 gene-disruptant strain. Thus, TTHB212 protein was identified as an anti-sigma(E) factor. These findings indicate that T. thermophilus HB8 has a regulatory system involving sigma(E) and anti-sigma(E) factors.
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