BsrG/SR4 from Bacillus subtilis- the first temperature-dependent type I toxin-antitoxin system

Summary Here, we describe bsrG/SR4, a novel type I toxin–antitoxin system from the SPβ prophage region of the Bacillus subtilis chromosome. The 294‐nucleotide bsrG RNA encodes a 38‐amino‐acid toxin, whereas SR4 is a 180‐nucleotide antisense RNA that acts as the antitoxin. Both genes overlap by 123 nucleotides. BsrG expression increases at the onset of stationary phase. The sr4 promoter is 6‐ to 10‐fold stronger than the bsrG promoter. Deletion of sr4 stabilizes bsrG mRNA and causes cell lysis on agar plates, which is due to the BsrG peptide and not the bsrG mRNA. SR4 overexpression could compensate cell lysis caused by overexpression of bsrG. SR4 interacts with the 3′ UTR of bsrG RNA, thereby promoting its degradation. RNase III cleaves the bsrG RNA/SR4 duplex at position 185 of bsrG RNA, but is not essential for the function of the toxin–antitoxin system. Endoribonuclease Y and 3′‐5′ exoribonuclease R participate in the degradation of both bsrG RNA and SR4, whereas PnpA processes three SR4 precursors to the mature RNA. A heat shock at 48°C results in faster degradation and, therefore, significantly decreased amounts of bsrG RNA.