COOPERATIVITY MUTANTS OF THE GAMMA-DELTA RESOLVASE IDENTIFY AN ESSENTIAL INTERDIMER INTERACTION
COOPERATIVITY MUTANTS OF THE GAMMA-DELTA RESOLVASE IDENTIFY AN ESSENTIAL INTERDIMER INTERACTION
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DOI：
10.1016/0092-8674(90)90428-h
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发表时间：
1990-12-21
期刊：
影响因子：
64.5
通讯作者：
GRINDLEY, NDF
中科院分区：
文献类型：
--
作者：
HUGHES, RE;HATFULL, GF;GRINDLEY, NDF
γ - β分解酶是一种转座子编码的位点特异性重组酶，催化γ - δ转位协整中间体的分解。重组反应涉及催化核蛋白复合物的形成，其结构由特定的蛋白质- dna和蛋白质-蛋白质相互作用决定。我们已经分离了许多分解突变体，并确定了四种无法介导重组所需的高阶蛋白质相互作用的亚类。这种突变表型的特点是无法催化重组，失去与res DNA的合作结合，以及无法诱导res内两个溶解性结合位点之间的DNA环出。通过合作突变体识别的氨基酸侧链聚集在蛋白质表面，介导晶体学四聚体中溶解性二聚体之间的相互作用。因此，我们已经确定了一个分解区域，该区域介导了重组突触中间体形成所必需的中间二聚体蛋白质-蛋白质相互作用。
Gamma-beta resolvase, a transposon-encoded site-specific recombinase, catalyzes the resolution of the cointegrate intermediate of gamma-delta transposition. The recombination reaction involves the formation of a catalytic nucleoprotein complex whose structure is determined by specific protein-DNA and protein-protein interactions. We have isolated many resolvase mutants and have identified four that are unable to mediate a subclass of higher order protein-protein interactions necessary for recombination. This mutant phenotype is characterized by an inability to catalyze recombination, a loss of cooperative binding to res DNA, and a failure to induce looping out of the DNA between two resolvase binding sites within res. The amino acid side chains identified by the cooperatively mutants cluster on a surface of the protein that mediates an interaction between resolvase dimers in a crystallographic tetramer. We have therefore identified a region of resolvase that mediates an interdimer protein-protein interaction necessary for the formation of the recombinogenic synaptic intermediate.