Domains formed within the N-terminal region of the quorum-sensing activator TraR are required for transcriptional activation and direct interaction with RpoA from agrobacterium

The Journal of Biological Chemistry
Y QinS K Farrand

Abstract

TraR, a quorum-sensing activator, induces transcription from its binding site, the tra-box, located upstream of Ti plasmid target promoters. TraR activated expression of a lacZ reporter in Escherichia coli only when RpoAAt from Agrobacterium tumefaciens was co-expressed. As assessed by gel retardation assays RpoAAt, but not RpoAEc, formed a ternary complex with TraR and a tra-box probe in vitro. TraR formed similar ternary complexes with alphaCTDAt but not with NTDAt, the C- and N-terminal segments of RpoAAt. As measured by surface plasmon resonance refractometry, TraR interacted directly with RpoAAt with an affinity about five times greater than that observed for its interaction with RpoAEc. The activator interacted with alphaCTDAt with kinetics and affinities similar to those of the full-sized -subunit. Positive control (PC) mutations at Asp-10 and Gly-123 of TraR did not affect DNA binding but greatly decreased the TraR-RpoAAt interaction. These two residues combine to form two patches on the activator, one of which may be involved in interaction with RpoA. When co-expressed, mutants of TraR with substitutions at Asp-10 complementing mutants with substitutions at Gly-123 for gene activation in an allele-specific manner. Co-e...Continue Reading

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Citations

Sep 6, 2006·Analytical and Bioanalytical Chemistry·William Nasser, Sylvie Reverchon
Nov 17, 2009·Foodborne Pathogens and Disease·Palmy R JesudhasanSuresh D Pillai
Mar 16, 2007·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Catharine E White, Stephen C Winans
Jul 15, 2009·Biotechnology and Applied Biochemistry·Sheetal RainaTajalli Keshavarz
Jul 4, 2015·Journal of Agricultural and Food Chemistry·Ram M UckooBhimanagouda S Patil
Jun 30, 2010·Communicative & Integrative Biology·Elise Haudecoeur, Denis Faure
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Dec 15, 2010·Chemical Reviews·Ann M StevensAlain Doutheau

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