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首页> 外文期刊>Biochemistry >Binding of MetJ Repressor to Specific and Nonspecific DNA and Effect of S-Adenosylmethionine on These Interactions
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Binding of MetJ Repressor to Specific and Nonspecific DNA and Effect of S-Adenosylmethionine on These Interactions

机译：MetJ阻遏物与特异性和非特异性DNA的结合以及S-腺苷甲硫氨酸对这些相互作用的影响

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摘要

We have used analytical ultracentrifugation to characterize the binding of the methioninenrepressor protein,MetJ, to synthetic oligonucleotides containing zero to five specific recognition sites, callednmetboxes. For all lengths of DNA studied,MetJ binds more tightly to repeats of the consensus sequence thannto naturally occurring metboxes, which exhibit a variable number of deviations from the consensus. Strongncooperative binding occurs only in the presence of two or more tandem metboxes, which facilitatenprotein-protein contacts between adjacent MetJ dimers, but weak affinity is detected even with DNAncontaining zero or one metbox. The affinity ofMetJ for all of the DNA sequences studied is enhanced by thenaddition of SAM, the known cofactor forMetJ in the cell. This effect extends to oligos containing zero or onenmetbox, both of which bind twoMetJ dimers. In the presence of a large excess concentration ofmetboxDNA,nthe effect of cooperativity is to favor populations of DNA oligos bound by two or more MetJ dimers rathernthan a stochastic redistribution of the repressor onto all available metboxes. These results illustrate thendynamic range of binding affinity and repressor assembly that MetJ can exhibit with DNA and the effect ofnthe corepressor SAM on binding to both specific and nonspecific DNA.
机译：我们已经使用分析超速离心法来表征蛋氨酸抑制蛋白MetJ与含有零至五个特定识别位点的合成寡核苷酸（称为nmetboxes）的结合。对于研究的所有长度的DNA，MetJ与共有序列的重复序列都比天然存在的metbox更紧密地结合，而天然存在的metbox表现出与共有序列不同的偏差。强合作性结合仅在两个或多个串联二元体存在下发生，这促进了相邻MetJ二聚体之间的蛋白质接触，但即使含有零或一个二元体的DNAn也检测到弱亲和力。然后通过添加SAM（已知的MetJ细胞辅助因子）来增强MetJ对所有研究的DNA序列的亲和力。这种作用扩展到包含零或onenmetbox的寡核苷酸，两者都结合两个MetJ二聚体。在大量过量的metboxDNA存在的情况下，协同作用的结果是有利于由两个或多个MetJ二聚体结合的DNA寡核苷酸群体，而不是阻遏物随机重新分布到所有可用的metbox上。这些结果说明了MetJ可以与DNA表现出的结合亲和力和阻遏物装配的动态范围，以及共抑制因子SAM对与特异性和非特异性DNA结合的影响。

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