在II类启动子转录激活过程中大肠杆菌CRP蛋白功能激活区域的取向
Orientation of functional activating regions in the Escherichia coli CRP protein during transcription activation at class II promoters.
作者信息
Williams R M, Rhodius V A, Bell A I, Kolb A, Busby S J
机构信息
School of Biochemistry, University of Birmingham, United Kingdom.
出版信息
Nucleic Acids Res. 1996 Mar 15;24(6):1112-8. doi: 10.1093/nar/24.6.1112.
Abstract
At class II CRP-dependent promoters the DNA site for CRP overlaps the DNA site for RNA polymerase, covering the -35 region. Transcription activation at class II CRP- dependent promoters requires a contact between an activating region in the upstream subunit of the bound CRP dimer and a contact site in the C-terminal domain of the alpha-subunit of RNA polymerase. Transcription activation is suppressed by amino acid substitutions in the activating region, but activation can be restored by second site substitutions at K52 or E96. These substitutions identify two separate regions on the surface of CRP that appear to be able to interact with RNA polymerase specifically at class II promoters. Using the method of 'oriented heterodimers' we show that these alternative activating regions are functional in the downstream subunit of the bound CRP dimer.
摘要
在II类CRP依赖性启动子中，CRP的DNA结合位点与RNA聚合酶的DNA结合位点重叠，覆盖了-35区域。II类CRP依赖性启动子的转录激活需要结合的CRP二聚体上游亚基中的激活区域与RNA聚合酶α亚基C末端结构域中的接触位点之间的接触。激活区域中的氨基酸替换会抑制转录激活，但K52或E96处的第二位点替换可恢复激活。这些替换确定了CRP表面上两个不同的区域，它们似乎能够在II类启动子处与RNA聚合酶特异性相互作用。使用“定向异二聚体”方法，我们表明这些替代激活区域在结合的CRP二聚体的下游亚基中起作用。
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