Transcriptional activation of a heat-shock gene, lonD, of Myxococcus xanthus by a two component histidine-aspartate phosphorelay system.

In vitro transcription of lonD, a heat-shock gene from Myxococcus xanthus, was stimulated in the presence of extract from heat-shocked cells. For this stimulation the upstream promoter region of lonD was found to be essential. Activation of lonD transcription was also observed when extract from non-heat-shocked cells was heat treated in vitro at 42 degrees C for 10 min. A DNA binding assay and footprinting analysis revealed that a factor(s) binds to the upstream region from -122 to -107 with respect to the transcription initiation site. This region was required for heat-shock induction of lonD expression both in vitro and in vivo. The lonD promoter-binding protein named HsfA was purified, and its gene was cloned. Analysis of the DNA sequence reveals that HsfA is a response regulator of the two-component system and shows high sequence similarity to the NtrC family or the enhancer-binding proteins. Upstream of hsfA, a gene encoding a histidine kinase was identified and named hsfB. HsfB was found to be autophosphorylated and able to phosphorylate HsfA. HsfA with HsfB activated in vitro transcription of lonD in a manner dependent on RNA polymerase containing SigA, the housekeeping sigma factor of M. xanthus.