Amino-terminal amino acids modulate sigma-factor DNA-binding activity.

Prokaryotic transcription initiation factor sigma is required for sequence-specific promoter recognition by RNA polymerase. Genetic studies have indicated that sigma itself interacts with DNA at the -10 and -35 promoter consensus sequences. Binding of Escherichia coli sigma 70 to DNA in vitro, however, can only be observed for truncated polypeptides lacking the amino-terminal amino acids. We have investigated the role of the amino terminus of E. coli sigma 70 in controlling DNA-binding ability. Deletion analysis indicates that amino acids within amino-terminal region 1.1 of sigma 70 inhibit DNA binding by the carboxy-terminal DNA-binding domains. Furthermore, inhibition of binding by the amino-terminal inhibitory domain of sigma 70 can be observed in trans. Likewise, the amino-terminal extensions of two alternative sigma-factors, E. coli sigma 32 and Bacillus subtilis sigma K, negatively affect the DNA binding activity of their carboxy-terminal domains. We propose that initiation of transcription is subject to modulation as a result of the composition and/or structure of the amino terminus of the sigma-subunit and that the sigma family of proteins belong to a larger class of intramolecularly regulated transcriptional effectors.