Identification and analysis of a principal sigma factor interacting protein sinA, essential for growth at high temperatures in a cyanobacterium synechococcus elongatus PCC 7942

Abstract

Proteins that bind to RNA polymerase (RNAP) sigma factors play important roles in various transcriptional regulations. In this study, we identified a candidate of the principal sigma factor interacting protein in cyanobacteria, named SinA, based on a previous comprehensive protein interaction study (Sato et al., 2007) and analyzed this in the cyanobacterium Synechococcus elongatus PCC 7942. SinA is highly conserved among cyanobacteria and a knock out mutant showed de-fective growth at a usually permissive high temperature (40°C). Because this observation sug-gested SinA involvement in heat-inducible transcriptional activation, we examined heat-inducible protein gene hspA expression after temperature upshifts. The second-step induction disappeared after 15 min in the sinA mutant. In vivo pull-down experiments demonstrated the interaction between SinA and the principal sigma factor RpoD1. This SinA-RpoD1 complex was associated with an RNAP core enzyme under growth temperatures, but was dissociated after a temperature upshift. Based on these results, we propose a function of SinA to facilitate the substitution of the principal sigma factor with alternative sigma factors under heat-stressed conditions.