首页> 外文期刊>Journal of Molecular Biology >SEQUENCE-SPECIFIC INTERACTIONS BETWEEN PROMOTER DNA AND THE RNA POLYMERASE SIGMA FACTOR E
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SEQUENCE-SPECIFIC INTERACTIONS BETWEEN PROMOTER DNA AND THE RNA POLYMERASE SIGMA FACTOR E

机译:启动子DNA与RNA聚合酶SIGMA因子E之间的序列特异性相互作用

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In order to determine which amino acyl residues in a secondary sigma factor govern its specificity of recognition at the -35 region of promoters, we examined the effects of amino acid substitutions in sigma(E) in Bacillus subtilis that made the sequence of its putative -35 recognition region more similar to another sigma factor in B. subtilis, sigma(K). We found that a single amino acid substitution at position 217 of sigma(E) resulted in a sigma factor that could direct transcription from sigma(K)-dependent promoters. Furthermore, we tested whether this amino acid substitution in sigma(E) had changed the specificity of interactions of the sigma with -35 region sequences by examining the activity of the mutant sigma(E) on derivatives of sigma(E)-dependent promoters that contained single base-pair substitutions. We found that this substitution in sigma(E) specifically suppressed the effect of a single base-pair substitution at position -31 in a sigma(E)-dependent promoter spoIIID. The amino acyl residue at another position (219) in sigma(E) affected the specificity of interaction with position -33 in spoIIID promoter. The amino acyl residues at the two positions in sigma(E), 217 and 219, that determine the specificity of interactions between the sigma and base-pairs in the -35 region of its cognate promoters (positions -33 and -31, respectively, in the spoIIID promoter) probably closely contact these base-pairs. (C) 1995 Academic Press Limited [References: 24]
机译:为了确定第二个sigma因子中的哪些氨基酰基残基控制其在启动子-35区的识别特异性,我们研究了枯草芽孢杆菌中sigma(E)中氨基酸取代的影响,该突变使推定的序列- 35个识别区域更类似于枯草芽孢杆菌中的另一个sigma因子sigma(K)。我们发现,在sigma(E)位置217的单个氨基酸取代会导致一个sigma因子,该因子可指导sigma(K)依赖型启动子的转录。此外,我们通过检查突变体sigma(E)对依赖于sigma(E)的启动子的衍生物的活性,测试了sigma(E)中的这种氨基酸取代是否已改变sigma与-35区域序列相互作用的特异性。包含单个碱基对取代。我们发现,在sigma(E)中的这种取代特别抑制了sigma(E)依赖性启动子spoIIID中位于-31位置的单个碱基对取代的影响。 Sigma(E)另一个位置(219)的氨基酰基残基影响与spoIIID启动子中-33位置相互作用的特异性。 σ(E)217和219中两个位置的氨基酰基残基决定了其同源启动子-35区域中σ和碱基对之间相互作用的特异性(分别为-33和-31)在SpoIIID启动子中)可能紧密接触这些碱基对。 (C)1995 Academic Press Limited [参考号:24]

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