ROLE OF THE ESCHERICHIA-COLI DNAK AND DNAJ HEAT-SHOCK PROTEINS IN THE INITIATION OF BACTERIOPHAGE-LAMBDA-DNA REPLICATION
ROLE OF THE ESCHERICHIA-COLI DNAK AND DNAJ HEAT-SHOCK PROTEINS IN THE INITIATION OF BACTERIOPHAGE-LAMBDA-DNA REPLICATION
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DOI：
10.1073/pnas.85.18.6632
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发表时间：
1988-09-01
影响因子：
11.1
通讯作者：
ZYLICZ, M
中科院分区：
文献类型：
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作者：
LIBEREK, K;GEORGOPOULOS, C;ZYLICZ, M
我们研究了两种大肠杆菌热休克蛋白，dna K和dna J基因产物，在体外启动λdv DNA复制过程中的作用。使用14C标记的.lambda。P蛋白，我们证明了DNAK和DNAJ热休克蛋白共同作用，释放.lambda。来自由lambda组成的前原粒复合体的P蛋白。复制的起点-.lambda。O-.Lambda。P-DNAB蛋白。在该反应中，可能由DNAK催化的ATP的水解是必需的。用突变的DNAK756蛋白替换DNAK蛋白可阻断lambda。P的释放。在DNAK和DNAJ作用后，在Sepharose4B上分离的前原粒复合体可以支持lambda。DV DNA复制不需要任何额外的预启动蛋白。使用DNAK-亲和层析，我们证明了这两个波长。O和.Lambda。P蛋白与DNAK蛋白结合。兰布达。P蛋白与DNAK蛋白以一种耐盐、疏水的方式相互作用，而ATP水解是必需的，以洗脱至少部分lambda。P蛋白从DNAK亲和层析柱中分离出来。原核生物DNAK和DNAJ热休克蛋白的作用机制与真核生物的DNAK类似物HSP70使用ATP来破坏疏水聚集体的假设一致[Pelham，H.R.B.(1986)Cell 46,959-961]。
We examined the role of two Escherichia coli heat shock proteins, the dnaK and dnaJ gene products, during the initiation of .lambda.dv DNA replication in vitro. Using 14C-labeled .lambda. P protein we showed that the DnaK and DnaJ heat shock proteins function together to release .lambda. P protein from the preprimosomal complex consisting of .lambda. origin of replication-.lambda. O-.lambda. P-DnaB protein. Hydrolysis of ATP, catalyzed presumably by DnaK, is required during this reaction. Substitution of DnaK protein with that of the mutant DnaK756 protein blocks .lambda. P release. After DnaK and DnaJ action, the preprimosomal complex, isolated on Sepharose 4B, can support .lambda. dv DNA replication without any additional prepriming proteins. Using DnaK-affinity chromatography we showed that both .lambda. O and .lambda. P proteins bind to DnaK protein. The .lambda. P protein interacts with DnaK protein in a salt-resistant, hydrophobic manner, and ATP hydrolysis is necessary to elute at least part of .lambda. P protein from the DnaK-affinity column. The proposed mechanism of action of the prokaryotic DnaK and DnaJ heat shock proteins agrees with the hypothesis that Hsp70, the DnaK analogue of eukaryotes, uses ATP to disrupt hydrophobic aggregates [Pelham, H. R. B. (1986) Cell 46, 959-961].