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UVM,一种大肠杆菌中紫外线诱导的不依赖RecA的诱变现象。

UVM, an ultraviolet-inducible RecA-independent mutagenic phenomenon in Escherichia coli.

作者信息

Palejwala V A, Pandya G A, Bhanot O S, Solomon J J, Murphy H S, Dunman P M, Humayun M Z

机构信息

Department of Microbiology and Molecular Genetics, UMD-New Jersey Medical School, Newark 07103-2714.

出版信息

J Biol Chem. 1994 Nov 4;269(44):27433-40.

PMID:7961656
Abstract

Most mutagenic DNA lesions are noninstructive in the sense that template instruction is either missing or inaccessible during DNA replication, leading to replication arrest. According to the SOS hypothesis, arrested replication induces the expression of SOS factors that force replication past stalled sites at the cost of mutagenesis. We have recently shown that prior UV irradiation of delta recA cells, in which the SOS pathway does not function, enhances mutagenesis at an ethenocytosine residue borne on a circular gapped duplex DNA vector, indicating the existence of an SOS-independent inducible mutagenic phenomenon termed UVM (UV modulation of mutagenesis). In the previous experiments, mutation fixation was expected to occur during gap-filling DNA synthesis. To test whether UVM is observable during normal replication by DNA polymerase III, we have examined mutagenesis at an epsilon C residue borne on M13 single-stranded DNA. By analyzing mutation frequency and specificity using a multiplex sequence assay, we now show that UVM is observable in UV-irradiated recA+, and in delta recA cells. These data indicate that UV irradiation induces a previously unrecognized mutagenic mechanism in Escherichia coli, and that this mechanism is manifested during gap-filling DNA synthesis as well as during normal DNA replication.

摘要

大多数诱变DNA损伤是非指令性的,因为在DNA复制过程中模板指令缺失或无法获取,从而导致复制停滞。根据SOS假说,停滞的复制会诱导SOS因子的表达,这些因子以诱变作用为代价促使复制越过停滞位点。我们最近发现,对SOS途径不起作用的δrecA细胞进行紫外线预照射,会增强携带于环状缺口双链DNA载体上的一个乙撑胞嘧啶残基处的诱变作用,这表明存在一种名为UVM(诱变作用的紫外线调制)的不依赖SOS的可诱导诱变现象。在之前的实验中,预计突变固定会在填补缺口的DNA合成过程中发生。为了测试在DNA聚合酶III进行正常复制期间是否能观察到UVM,我们检测了M13单链DNA上携带的一个εC残基处的诱变情况。通过使用多重序列分析法分析突变频率和特异性,我们现在表明在紫外线照射的recA+细胞以及δrecA细胞中都能观察到UVM。这些数据表明紫外线照射在大肠杆菌中诱导了一种先前未被认识的诱变机制,并且这种机制在填补缺口的DNA合成以及正常DNA复制过程中都会表现出来。

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