Mukaihara T, Enomoto M
Department of Biology, Faculty of Science, Okayama University, Japan.
Genetics. 1997 Mar;145(3):563-72. doi: 10.1093/genetics/145.3.563.
Deletion formation between the 5'-mostly homologous sequences and between the 3'-homeologous sequences of the two Salmonella typhimurium flagellin genes was examined using plasmid-based deletion-detection systems in various Escherichia coli genetic backgrounds. Deletions in plasmid pLC103 occur between the 5' sequences, but not between the 3' sequences, in both RecA-independent and RecA-dependent ways. Because the former is predominant, deletion formation in a recA background depends on the length of homologous sequences between the two genes. Deletion rates were enhanced 30- to 50-fold by the mismatch repair defects, mutS, mutL and uvrD, and 250-fold by the ssb-3 allele, but the effect of the mismatch defects was canceled by the delta recA allele. Rates of the deletion between the 3' sequences in plasmid pLC107 were enhanced 17- to 130-fold by ssb alleles, but not by other alleles. For deletions in pLC107, 96% of the endpoints in the recA+ background and 88% in delta recA were in the two hot spots of the 60- and 33-nucleotide (nt) homologous sequences, whereas in the ssb-3 background > 50% of the endpoints were in four- to 14-nt direct repeats dispersed in the entire 3' sequences. The deletion formation between the homeologous sequences in RecA-independent but depends on the length of consecutive homologies. The mutant ssb allele lowers this dependency and results in the increase in deletion rates. Roles of mutant SSB are discussed with relation to misalignment in replication slippage.
利用基于质粒的缺失检测系统,在各种大肠杆菌遗传背景下,研究了两个鼠伤寒沙门氏菌鞭毛蛋白基因5'端大部分同源序列之间以及3'端同源序列之间的缺失形成情况。质粒pLC103中的缺失发生在5'序列之间,而非3'序列之间,且存在RecA非依赖型和RecA依赖型两种方式。由于前者占主导,recA背景下的缺失形成取决于两个基因之间同源序列的长度。错配修复缺陷mutS、mutL和uvrD使缺失率提高了30至50倍,而ssb - 3等位基因使其提高了250倍,但错配缺陷的影响被ΔrecA等位基因抵消。质粒pLC107中3'序列之间的缺失率被ssb等位基因提高了至17至130倍,但其他等位基因无此作用。对于pLC107中的缺失,recA +背景下96%的断点和ΔrecA背景下88%的断点位于60和33个核苷酸(nt)同源序列的两个热点区域,而在ssb - 3背景下,超过50%的断点位于分散在整个3'序列中的4至14 nt直接重复序列中。同源序列之间的缺失形成是RecA非依赖型的,但取决于连续同源性的长度。突变的ssb等位基因降低了这种依赖性并导致缺失率增加。结合复制滑动中的错配情况讨论了突变型单链结合蛋白(SSB)的作用