Modulation of T4 gene 32 protein DNA binding activity by the recombination mediator protein UvsY.

Modulation of T4 gene 32 protein DNA binding activity by the recombination mediator protein UvsY.
复制标题

重组介体蛋白 UvsY 对 T4 基因 32 蛋白 DNA 结合活性的调节。

DOI:
10.1016/j.jmb.2008.05.039
复制
发表时间:
2008
影响因子:
5.6
通讯作者:
Williams,MarkC
Williams,MarkC
中科院分区:
生物学2区
文献类型:
--
作者:
Pant,Kiran;Shokri,Leila;Karpel,RichardL;Morrical,ScottW;Williams,MarkC

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噬菌体T4 UvsY是一种重组中介蛋白,可促进uvx - ssdna突触前丝的组装。UvsY帮助UvsX从ssDNA中取代T4基因32蛋白(gp32),这是突触前丝正确形成所必需的反应。在这里,我们使用DNA拉伸来检测在gp32和gp32 c端截断(*I)存在和不存在的情况下UvsY与单个DNA分子的相互作用,并表明在这两种情况下,UvsY都能够破坏gp32- ssdna相互作用的稳定性。在这些实验中,UvsY与dsDNA的结合比ssDNA更强,因为它无法在高强度下包裹ssDNA。为了支持这一假设,我们表明,拉伸DNA暴露于乙二醛产生的ssDNA被UvsY强烈包裹,但包裹只发生在低力下。我们的研究结果表明,在没有其他蛋白质的情况下,UvsY与拉伸的DNA有很强的相互作用。在gp32和*I存在的情况下,UvsY能够强烈破坏gp32- dna复合物的稳定,从而促进ssDNA的包裹,从而在UvsX存在的情况下为突触前丝组装准备ssDNA。因此,UvsY通过将刚性gp32-DNA细丝转化为可被UvsX强结合的结构来介导UvsX与ssDNA的结合。
Bacteriophage T4 UvsY is a recombination mediator protein that promotes assembly of the UvsX-ssDNA presynaptic filament. UvsY helps UvsX to displace T4 gene 32 protein (gp32) from ssDNA, a reaction necessary for proper formation of the presynaptic filament. Here we use DNA stretching to examine UvsY interactions with single DNA molecules in the presence and absence of gp32 and a gp32 C-terminal truncation (*I), and show that in both cases UvsY is able to destabilize gp32-ssDNA interactions. In these experiments UvsY binds more strongly to dsDNA than ssDNA due to its inability to wrap ssDNA at high forces. To support this hypothesis, we show that ssDNA created by exposure of stretched DNA to glyoxal is strongly wrapped by UvsY, but wrapping occurs only at low forces. Our results demonstrate that UvsY interacts strongly with stretched DNA in the absence of other proteins. In the presence of gp32 and *I, UvsY is capable of strongly destabilizing gp32-DNA complexes in order to facilitate ssDNA wrapping, which in turn prepares the ssDNA for presynaptic filament assembly in the presence of UvsX. Thus, UvsY mediates UvsX binding to ssDNA by converting rigid gp32-DNA filaments into a structure that can be strongly bound by UvsX.
蛋白质-蛋白质相互作用在 T4 同源重组突触前丝组装中的作用。
DOI: --
发表时间: 1993
期刊: The Journal of biological chemistry
影响因子: --
作者:
Jiang,H;Giedroc,D;Kodadek,T
通讯作者: Kodadek,T
DOI: 10.1021/bi9817055
发表时间: 1999-01
期刊: Biochemistry
影响因子: 2.9
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DOI: 10.1016/s0006-3495(01)76066-3
发表时间: 2001-02-01
影响因子: 3.4
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Williams, MC;Wenner, JR;Bloomfield, VA
通讯作者: Bloomfield, VA
T4 噬菌体基因 32:DNA 复制和重组中的结构蛋白
DOI: --
发表时间: 1970
期刊: Nature
影响因子: 64.8
作者:
B. Alberts;L. Frey
通讯作者: L. Frey
DOI: 10.1016/s0006-3495(01)76163-2
发表时间: 2001-04-01
影响因子: 3.4
作者:
Williams, MC;Wenner, JR;Bloomfield, VA
通讯作者: Bloomfield, VA