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DNA聚合酶III全酶的β亚基在大肠杆菌经紫外线照射或萘啶酸处理后被诱导产生。

Beta subunit of DNA polymerase III holoenzyme is induced upon ultraviolet irradiation or nalidixic acid treatment of Escherichia coli.

作者信息

Tadmor Y, Bergstein M, Skaliter R, Shwartz H, Livneh Z

机构信息

Department of Biochemistry, Weizmann Institute of Science, Rehovot, Israel.

出版信息

Mutat Res. 1994 Jul 1;308(1):53-64. doi: 10.1016/0027-5107(94)90198-8.

Abstract

Exposure of Escherichia coli to UV irradiation or nalidixic acid, which induce both the SOS and heat shock responses, led to a 3-4-fold increase in the amount of the beta subunit of DNA polymerase III holoenzyme, as assayed by Western blot analysis using anti-beta antibodies. Such an induction was observed also in a delta rpoH mutant lacking the heat shock-specific sigma 32 subunit of RNA polymerase, but it was not observed in recA13 or lexA3 mutants, in which the SOS response cannot be induced. Mapping of transcription initiation sites of the dnaN gene, encoding the beta subunit, using the S1 nuclease protection assay showed essentially no induction of transcription upon UV irradiation, indicating that induction is regulated primarily at the post-transcriptional level. Analysis of translational gene fusions of the dnaN gene, encoding the beta subunit, to the lacZ reporter gene showed induction of beta-galactosidase activity upon UV irradiation of cells harboring the fusion plasmids. Elimination of a 5' flanking DNA sequence in which the dnaN promoters P1 and P2 were located, did not affect the UV inducibility of the gene fusions. Thus, element(s) present from P3 downstream were sufficient for the UV induction. The induction of the dnaN-lacZ gene fusions was dependent on the recA and lexA gene products, but not on the rpoH gene product, in agreement with the immunoblot analysis. The dependence of dnaN induction on the SOS regulators was not mediated via classical repression by the LexA repressor, since the dnaN promoter does not contain a sequence homologous to the LexA binding site, and dnaN mRNA was not inducible by UV light. This suggests that SOS control may be imposed indirectly, by a post-transcriptional mechanism. The increased amount of the beta subunit is needed, most likely, for increased replication and repair activities in cells which have been exposed to UV radiation.

摘要

将大肠杆菌暴露于紫外线照射或萘啶酸下,这两者都会诱导SOS和热休克反应,通过使用抗β抗体的蛋白质印迹分析测定,DNA聚合酶III全酶的β亚基量增加了3至4倍。在缺乏RNA聚合酶热休克特异性σ32亚基的ΔrpoH突变体中也观察到了这种诱导,但在recA13或lexA3突变体中未观察到,在这些突变体中无法诱导SOS反应。使用S1核酸酶保护试验对编码β亚基的dnaN基因的转录起始位点进行定位,结果表明紫外线照射后基本上没有转录诱导,这表明诱导主要在转录后水平受到调控。对编码β亚基的dnaN基因与lacZ报告基因的翻译基因融合体进行分析,结果表明,对携带融合质粒的细胞进行紫外线照射后,β-半乳糖苷酶活性会被诱导。去除dnaN启动子P1和P2所在的5'侧翼DNA序列,并不影响基因融合体的紫外线诱导性。因此,P3下游存在的元件足以实现紫外线诱导。dnaN-lacZ基因融合体的诱导依赖于recA和lexA基因产物,但不依赖于rpoH基因产物,这与免疫印迹分析结果一致。dnaN诱导对SOS调节因子的依赖性不是通过LexA阻遏物的经典抑制介导的,因为dnaN启动子不包含与LexA结合位点同源的序列,并且dnaN mRNA不能被紫外线诱导。这表明SOS控制可能是通过转录后机制间接施加的。增加β亚基的量很可能是为了增加暴露于紫外线辐射的细胞中的复制和修复活性。

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