Ramírez-Guadiana Fernando H, Barajas-Ornelas Rocío Del Carmen, Corona-Bautista Saúl U, Setlow Peter, Pedraza-Reyes Mario
Department of Biology, Division of Natural and Exact Sciences, University of Guanajuato, Guanajuato, Gto. 36050, México.
Department of Molecular Biology and Biophysics, UConn Health, 06030-3305, Farmington, Connecticut, United States of America.
PLoS One. 2016 Mar 1;11(3):e0150348. doi: 10.1371/journal.pone.0150348. eCollection 2016.
The expression of and role played by RecA in protecting sporulating cells of Bacillus subtilis from DNA damage has been determined. Results showed that the DNA-alkylating agent Mitomycin-C (M-C) activated expression of a PrecA-gfpmut3a fusion in both sporulating cells' mother cell and forespore compartments. The expression levels of a recA-lacZ fusion were significantly lower in sporulating than in growing cells. However, M-C induced levels of ß-galactosidase from a recA-lacZ fusion ~6- and 3-fold in the mother cell and forespore compartments of B. subtilis sporangia, respectively. Disruption of recA slowed sporulation and sensitized sporulating cells to M-C and UV-C radiation, and the M-C and UV-C sensitivity of sporangia lacking the transcriptional repair-coupling factor Mfd was significantly increased by loss of RecA. We postulate that when DNA damage is encountered during sporulation, RecA activates the SOS response thus providing sporangia with the repair machinery to process DNA lesions that may compromise the spatio-temporal expression of genes that are essential for efficient spore formation.
已确定RecA在保护枯草芽孢杆菌的芽孢形成细胞免受DNA损伤方面的表达及所起的作用。结果表明,DNA烷化剂丝裂霉素-C(M-C)激活了芽孢形成细胞的母细胞和前芽孢区室中PrecA-gfpmut3a融合蛋白的表达。recA-lacZ融合蛋白在芽孢形成细胞中的表达水平显著低于生长细胞。然而,M-C诱导枯草芽孢杆菌孢子囊母细胞和前芽孢区室中recA-lacZ融合蛋白产生的β-半乳糖苷酶水平分别约为原来的6倍和3倍。recA的缺失减缓了芽孢形成,并使芽孢形成细胞对M-C和UV-C辐射敏感,而缺乏转录修复偶联因子Mfd的孢子囊对M-C和UV-C的敏感性因RecA的缺失而显著增加。我们推测,在芽孢形成过程中遇到DNA损伤时,RecA会激活SOS反应,从而为孢子囊提供修复机制,以处理可能损害高效孢子形成所必需基因的时空表达的DNA损伤。