Mashimo Kazumi, Kawata Masakado, Yamamoto Kazuo
Department of Biomolecular Sciences, Graduate School of Life Sciences, Tohoku University, Sendai 980-8577, Japan.
Mutagenesis. 2003 Jul;18(4):355-63. doi: 10.1093/mutage/geg004.
The endogenous tonB gene of Escherichia coli was used as a target for spontaneous deletion mutations which were isolated from recJ(-) and recQ(-) cells. Large deletions, due to simultaneous mutations of the trp operon, were also isolated. The rates of tonB mutation were 2.77 x 10(-8), 4.13 x 10(-8) and 5.00 x 10(-8) for rec(+), recJ(-) and recQ(-) cells, respectively. We analyzed 94 and 99 tonB mutants from the recJ(-) and recQ(-) cells, respectively, by sequencing. We found that IS insertion dominated, followed by base substitutions, frameshifts and deletions in both recJ(-) and recQ(-) strains. We then analyzed 55 tonB-trp deletions, ranging in size from 5907 to 20,832 bp, from the recJ(-) strains and 47 tonB-trp deletions, ranging in size from 4,959 to 16,390 bp from the recQ(-) strains. About one-third of tonB-trp deletions from both the recJ(-) and the recQ(-) cells were found to have occurred between short sequence repeats at the deletion termini. About one-third of tonB-trp deletions from both mutants showed 2-4 bp repeats in the immediate vicinity of the endpoints, which appeared to indicate no clear association with deletion. The remaining one-third of tonB-trp deletions had no homology at the endpoint. These results were similar to those for the rec(+) cells. Hanada and colleagues demonstrated that structually similar rearrangements arising during lambda bio phage formation (illegitimate recombination) increased in the recQ(-) strain. To explain this discrepancy, we interpreted as distinctive the mechanism for rearrangement during transducing phage formation which is recQ-dependent and that for deletions formed in chromosomes which is recQ-independent.
大肠杆菌的内源性tonB基因被用作自发缺失突变的靶点,这些突变是从recJ(-)和recQ(-)细胞中分离出来的。同时,由于色氨酸操纵子的同时突变,也分离出了大的缺失突变。rec(+)、recJ(-)和recQ(-)细胞的tonB突变率分别为2.77×10(-8)、4.13×10(-8)和5.00×10(-8)。我们分别对recJ(-)和recQ(-)细胞中的94个和99个tonB突变体进行了测序分析。我们发现,在recJ(-)和recQ(-)菌株中,IS插入占主导地位,其次是碱基替换、移码和缺失。然后,我们分析了recJ(-)菌株中55个大小从5907到20832 bp的tonB-trp缺失以及recQ(-)菌株中47个大小从4959到16390 bp的tonB-trp缺失。发现recJ(-)和recQ(-)细胞中约三分之一的tonB-trp缺失发生在缺失末端的短序列重复之间。两个突变体中约三分之一的tonB-trp缺失在端点附近显示出2-4 bp的重复,这似乎表明与缺失没有明显关联。其余三分之一的tonB-trp缺失在端点处没有同源性。这些结果与rec(+)细胞的结果相似。花田及其同事证明,在λ生物噬菌体形成过程中出现的结构相似的重排(非法重组)在recQ(-)菌株中增加。为了解释这种差异,我们将转导噬菌体形成过程中依赖recQ的重排机制与染色体中形成的不依赖recQ的缺失机制解释为不同的机制。