Ludanyi Monika, Blanchard Laurence, Dulermo Rémi, Brandelet Géraldine, Bellanger Laurent, Pignol David, Lemaire David, de Groot Arjan
CEA, DSV, IBEB, Lab Bioenerget Cellulaire, Saint-Paul-lez-Durance, F-13108, France; CNRS, UMR 7265 Biol Veget & Microbiol Environ, Saint-Paul-lez-Durance, F-13108, France; Aix-Marseille Université, Saint-Paul-lez-Durance, F-13108, France.
Mol Microbiol. 2014 Oct;94(2):434-49. doi: 10.1111/mmi.12774. Epub 2014 Sep 18.
Deinococcus bacteria are famous for their extreme radiation tolerance. The IrrE protein was shown to be essential for radiation tolerance and, in an unelucidated manner, for induction of a number of genes in response to radiation, including recA and other DNA repair genes. Earlier studies indicated that IrrE could be a zinc peptidase, but proteolytic activity was not demonstrated. Here, using several in vivo and in vitro experiments, IrrE from Deinococcus deserti was found to interact with DdrO, a predicted regulator encoded by a radiation-induced gene that is, like irrE, highly conserved in Deinococcus. Moreover, IrrE was found to cleave DdrO in vitro and when the proteins were coexpressed in Escherichia coli. This cleavage was not observed in the presence of metal chelator EDTA or when IrrE contains a mutation in the conserved active-site motif of metallopeptidases. In D. deserti, IrrE-dependent cleavage of DdrO was observed after exposure to radiation. Furthermore, DdrO-dependent repression of the promoter of a radiation-induced gene was shown. These results demonstrate that IrrE is a metalloprotease and we propose that IrrE-mediated cleavage inactivates repressor protein DdrO, leading to transcriptional induction of various genes required for repair and survival after exposure of Deinococcus to radiation.
嗜热栖热菌属细菌以其极强的辐射耐受性而闻名。IrrE蛋白被证明是辐射耐受性所必需的,并且以一种未阐明的方式,参与响应辐射诱导多个基因,包括recA和其他DNA修复基因。早期研究表明IrrE可能是一种锌肽酶,但未证实其蛋白水解活性。在这里,通过一些体内和体外实验,发现来自沙漠嗜热栖热菌的IrrE与DdrO相互作用,DdrO是一种由辐射诱导基因编码的预测调节因子,与irrE一样,在嗜热栖热菌属中高度保守。此外,发现IrrE在体外以及当这些蛋白在大肠杆菌中共表达时可切割DdrO。在存在金属螯合剂EDTA的情况下或当IrrE在金属肽酶的保守活性位点基序中发生突变时,未观察到这种切割。在沙漠嗜热栖热菌中,暴露于辐射后观察到IrrE依赖的DdrO切割。此外,还表明了DdrO对辐射诱导基因启动子的抑制作用。这些结果表明IrrE是一种金属蛋白酶,我们提出IrrE介导的切割使阻遏蛋白DdrO失活,导致嗜热栖热菌暴露于辐射后修复和存活所需的各种基因的转录诱导。