Regulation of late-acting operons by three transcription factors and a CRISPR-Cas component duringMyxococcus xanthusdevelopment

Abstract

SummaryUpon starvation rod-shapedMyxococcus xanthusbacteria form mounds and then differentiate into round stress-resistant spores. Little is known about the regulation of late-acting operons important for spore formation. C-signaling has been proposed to activate FruA, which binds DNA cooperatively with MrpC to increase transcription of many genes. We report that this model can explain regulation of thefadIJoperon involved in spore metabolism, but not that of the spore coat biogenesis operonsexoA-I,exoL-P, andnfsA-H. Rather, a mutation infruAincreased the transcript levels from these operons early in development, suggesting negative regulation by FruA initially, and a mutation inmrpCaffected transcript levels from each operon differently. FruA bound to all four promoter regionsin vitro, but strikingly each promoter region was unique in terms of whether or not MrpC and the DNA-binding domain of Nla6 bound, and in terms of cooperative binding. Furthermore, the DevI component of a CRISPR-Cas system is a negative regulator of all four operons, based on transcript measurements. Our results demonstrate complex regulation of sporulation genes by three transcription factors and a CRISPR-Cas component, which we propose thwarts viral intrusion while making spores suited to withstand starvation and environmental insults.