Departments of Molecular and Human Genetics, Biochemistry and Molecular Biology, Molecular Virology and Microbiology and the Dan L Duncan Cancer Center, Baylor College of Medicine, Houston, TX 77030-3411, United States.
DNA Repair (Amst). 2013 Nov;12(11):993-9. doi: 10.1016/j.dnarep.2013.08.011. Epub 2013 Sep 25.
Thymineless death (TLD) is the rapid loss of colony-forming ability in bacterial, yeast and human cells starved for thymine, and is the mechanism of action of common chemotherapeutic drugs. In Escherichia coli, significant loss of viability during TLD requires the SOS replication-stress/DNA-damage response, specifically its role in inducing the inhibitor of cell division, SulA. An independent RecQ- and RecJ-dependent TLD pathway accounts for a similarly large additional component of TLD, and a third SOS- and RecQ/J-independent TLD pathway has also been observed. Although two groups have implicated the SOS-response in TLD, an SOS-deficient mutant strain from an earlier study was found to be sensitive to thymine deprivation. We performed whole-genome resequencing on that SOS-deficient strain and find that, compared with the SOS-proficient control strain, it contains five mutations in addition to the SOS-blocking lexA(Ind(-)) mutation. One of the additional mutations, csrA, confers TLD sensitivity specifically in SOS-defective strains. We find that CsrA, a carbon storage regulator, reduces TLD in SOS- or SulA-defective cells, and that the increased TLD that occurs in csrA(-) SOS-defective cells is dependent on RecQ. We consider a hypothesis in which the modulation of nucleotide pools by CsrA might inhibit TLD specifically in SOS-deficient (SulA-deficient) cells.
胸腺嘧啶缺乏性死亡(TLD)是细菌、酵母和人类细胞在缺乏胸腺嘧啶时迅速丧失集落形成能力的机制,也是常见化疗药物的作用机制。在大肠杆菌中,TLD 期间显著丧失活力需要 SOS 复制应激/DNA 损伤反应,特别是其在诱导细胞分裂抑制剂 SulA 中的作用。独立的 RecQ 和 RecJ 依赖性 TLD 途径占 TLD 的一个类似大的额外组成部分,并且还观察到第三个 SOS 和 RecQ/J 独立的 TLD 途径。尽管有两个小组将 SOS 反应与 TLD 联系起来,但早期研究中的 SOS 缺陷突变株被发现对胸腺嘧啶剥夺敏感。我们对该 SOS 缺陷株进行了全基因组重测序,发现与 SOS 功能正常的对照株相比,除了 SOS 阻断 lexA(Ind(-))突变外,它还包含五个突变。除了 SOS 阻断 lexA(Ind(-))突变外,其中一个额外的突变 csrA 特异性赋予 TLD 敏感性。我们发现 CsrA,一种碳储存调节剂,可降低 SOS 或 SulA 缺陷细胞中的 TLD,并且在 csrA(-)SOS 缺陷细胞中发生的增加的 TLD 依赖于 RecQ。我们考虑了一种假设,即 CsrA 对核苷酸池的调节可能特异性地抑制 SOS 缺陷(SulA 缺陷)细胞中的 TLD。