A Novel Factor Required for the Assembly of the DnaK and DnaJ Chaperones of Thermus thermophilus*

A Novel Factor Required for the Assembly of the DnaK and DnaJ Chaperones of Thermus thermophilus*
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嗜热栖热菌 DnaK 和 DnaJ 分子伴侣组装所需的新因子*

DOI:
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发表时间:
1996
影响因子:
4.8
通讯作者:
Masasuke Yoshida
Masasuke Yoshida
中科院分区:
生物学2区
文献类型:
--
作者:
K. Motohashi;M. Yohda;I. Endo;Masasuke Yoshida

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我们曾报道过从嗜热栖热菌中分离到T.DnaK·DnaJ分子伴侣复合物。在这里,我们表明,一个新的因素是必要的组装T.DnaK和T.DnaJ到复杂的。结果表明,在T.嗜热菌含有dnaK-grpE-dnaJ-orf 4-clpB五个基因。有趣的是,T.DnaJ缺乏整个“富含半胱氨酸的区域”,该区域被认为是结合未折叠蛋白质所必需的。orf 4基因编码一种新的78个氨基酸的蛋白质。奇怪的是,在大肠杆菌中表达的T.DnaK和T.DnaJ没有形成复合物。对T.DnaK·DnaJ复合物的仔细重新检查显示,复合物中存在一种小蛋白,它原来是orf 4的产物。如预期的那样,dnaK-dnaJ-orf 4三个基因的表达导致在E.大肠杆菌中,这是无法区分从真正的复合物的能力,与核苷酸和变性蛋白质。orf 4的产物也是体外重组复合物所必需的,被命名为T.DnaK·DnaJ组装因子A(T.DnaK·DnaJ assembly factor A)。该复合物包含T.DnaK、T.DnaJ和T. DafA各三个拷贝。尽管在数据库中还没有发现T.DafA的明确同源物,但这一发现提出了一种可能性,即其他生物中DnaK和DnaJ分子伴侣之间的相互作用也是由一种尚未被注意到的小蛋白介导的。
We previously reported the isolation of T.DnaK·DnaJ chaperone complex from Thermus thermophilus. Here, we show that a novel factor is necessary for the assembly of T.DnaK and T.DnaJ into the complex. A dnaK gene cluster of T. thermophilus contained five genes, dnaK-grpE-dnaJ-orf4-clpB. Interestingly, T.DnaJ lacks the whole “cysteine-rich region” that has been postulated to be necessary to bind unfolded proteins. The orf4 gene encodes a novel 78-amino acid protein. Curiously, T.DnaK and T.DnaJ expressed in Escherichia coli did not form the complex. Careful reexamination of the T.DnaK·DnaJ complex revealed the presence of a small protein in the complex, which turned out to be a product of orf4. As expected, expression of three genes, dnaK-dnaJ-orf4, resulted in production of a T.DnaK·DnaJ complex in E. coli that was indistinguishable from the authentic complex in its ability to interact with nucleotide and denatured protein. The product of orf4 was also required for in vitro reconstitution of the complex and named T.DafA (T.DnaK·DnaJ assembly factor A). The complex comprises three copies each of T.DnaK, T.DnaJ, and T.DafA. Even though a definite homolog of T.DafA has not been found in the data base, this finding raises a possibility that interaction between DnaK and DnaJ chaperones in other organisms is also mediated by a small protein yet unnoticed.
DOI: 10.1016/s0021-9258(18)53470-0
发表时间: 1993-03
期刊: The Journal of biological chemistry
影响因子: --
作者:
J. Osipiuk;C. Georgopoulos;M. Żylicz
通讯作者: J. Osipiuk;C. Georgopoulos;M. Żylicz
DOI: 10.1016/s0021-9258(18)45398-7
发表时间: 1987-12
期刊: The Journal of biological chemistry
影响因子: --
作者:
M. Żylicz;D. Ang;Costa Georgopoulos
通讯作者: M. Żylicz;D. Ang;Costa Georgopoulos
DOI: --
发表时间: 1993
期刊: The Journal of biological chemistry
影响因子: --
作者:
Ziemienowicz,A;Skowyra,D;Zeilstra-Ryalls,J;Fayet,O;Georgopoulos,C;Zylicz,M
通讯作者: Zylicz,M
DOI: 10.1073/pnas.91.22.10345
发表时间: 1994-10-25
影响因子: 11.1
作者:
SZABO, A;LANGER, T;HARTL, FU
通讯作者: HARTL, FU
DOI: --
发表时间: 1986
期刊: The Journal of biological chemistry
影响因子: --
作者:
Bardwell,JC;Tilly,K;Craig,E;King,J;Zylicz,M;Georgopoulos,C
通讯作者: Georgopoulos,C