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GA结合蛋白的α亚基和β亚基在溶液中形成稳定的异二聚体。异源四聚体复合物组装的修订模型。

The alpha and beta subunits of the GA-binding protein form a stable heterodimer in solution. Revised model of heterotetrameric complex assembly.

作者信息

Chinenov Y, Henzl M, Martin M E

机构信息

Department of Biochemistry, University of Missouri at Columbia, Columbia, Missouri 65212, USA.

出版信息

J Biol Chem. 2000 Mar 17;275(11):7749-56. doi: 10.1074/jbc.275.11.7749.

DOI:10.1074/jbc.275.11.7749
PMID:10713087
Abstract

We have studied the assembly of GA-binding protein (GABP) in solution and established the role of DNA in the assembly of the transcriptionally active GABPalpha(2)beta(2) heterotetrameric complex. GABP binds DNA containing a single PEA3/Ets-binding site (PEA3/EBS) exclusively as the alphabeta heterodimer complex, but readily binds as the GABPalpha(2)beta(2) heterotetramer complex on DNA containing two PEA3/EBSs. Positioning of the PEA3/EBSs on the same face of the DNA helix stabilizes heterotetramer complex binding. These observations suggest that GABPalphabeta heterodimers are the predominant molecular species in solution and that DNA containing two PEA3/EBSs promotes formation of the GABPalpha(2)beta(2) heterotetrameric complex. We analyzed the assembly of GABPalpha(2)beta(2) heteromeric complexes in solution by analytical ultracentrifugation. GABPalpha exists as a monomer in solution while GABPbeta exists in a monomer-dimer equilibrium (K(d) = 1.8 +/- 0.27 microM). In equimolar mixtures of the two subunits, GABPalpha and GABPbeta formed a stable heterodimer, with no heterotetramer complex detected. Thus, GABP exists in solution as the heterodimer previously shown to be a weak transcriptional activator. Assembly of the transcriptionally active GABPalpha(2)beta(2) heterotetramer complex requires the presence of specific DNA containing at least two PEA3/EBSs.

摘要

我们研究了GA结合蛋白（GABP）在溶液中的组装，并确定了DNA在转录活性GABPalpha(2)beta(2)异源四聚体复合物组装中的作用。GABP仅作为αβ异源二聚体复合物特异性结合含有单个PEA3/Ets结合位点（PEA3/EBS）的DNA，但在含有两个PEA3/EBS的DNA上很容易作为GABPalpha(2)beta(2)异源四聚体复合物结合。PEA3/EBS在DNA螺旋同一面上的定位稳定了异源四聚体复合物的结合。这些观察结果表明，GABPalphabeta异源二聚体是溶液中的主要分子形式，并且含有两个PEA3/EBS的DNA促进了GABPalpha(2)beta(2)异源四聚体复合物的形成。我们通过分析超速离心分析了溶液中GABPalpha(2)beta(2)异聚体复合物的组装。GABPalpha在溶液中以单体形式存在，而GABPbeta以单体-二聚体平衡形式存在（K(d)=1.8±0.27 microM）。在两个亚基的等摩尔混合物中，GABPalpha和GABPbeta形成了稳定的异源二聚体，未检测到异源四聚体复合物。因此，GABP在溶液中以异源二聚体形式存在，先前已证明其为弱转录激活剂。转录活性GABPalpha(2)beta(2)异源四聚体复合物的组装需要存在至少含有两个PEA3/EBS的特定DNA。

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