Alternative sigma factors play a key role in the physiology of Bacillus anthracis by regulating the transcription of the appropriate genes required for adaptation and survival. Under specific conditions, alternative sigma factors activate transcription by binding to the promoter of the genes relevant to the condition and initiate synthesis of RNA. Here we report that the transcription of predicted sigI gene in B. anthracis, BAS3231, is induced by elevated temperatures and involved in the regulation of virulence gene expression. We show that BAS3231 is required for cell viability at elevated temperatures. We have also demonstrated that mutation in the BAS3231 gene results in a decrease in virulence gene expression. Our study provides new insight into the role of alternative sigma factors in B. anthracis.