Bacterial protein translocation requires only one copy of the SecY complex in vivo.

Bacterial protein translocation requires only one copy of the SecY complex in vivo.
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DOI:
10.1083/jcb.201205140
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发表时间:
2012-09-03
期刊:
The Journal of cell biology
影响因子:
--
通讯作者:
Rapoport TA
Rapoport TA
中科院分区:
其他
文献类型:
--
作者:
Park E;Rapoport TA

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SecY在共翻译和翻译后易位过程中的寡聚状态的体内探测揭示了该过程不需要寡聚化。蛋白质在细菌中跨质膜的运输需要由SecY复合物形成的通道,该通道在共翻译易位中与翻译核糖体或在翻译后易位中与SecA ATP酶合作。易位是否需要SecY复合物的寡聚体是一个重要但有争议的问题:它决定了通道的大小,如何防止小分子的渗透,以及通道如何与核糖体和SecA相互作用。在这里,我们探测在体内的低聚状态的SecY交联,使用定义的共同和翻译后易位中间体在完整的大肠杆菌细胞。我们发现,nontranslocating SecY关联瞬时通过不同的相互作用表面与膜内的其他SecY分子。这些相互作用显着减少时,易位多肽插入SecY通道的共同或post-actionally。突变消除SecY分子之间的相互作用仍然支持E。杆菌这些结果表明,单个SecY分子足以进行蛋白质易位。
In vivo probing of the oligomeric state of SecY during co- and post-translational translocation reveals that oligomerization is not required for this process. The transport of proteins across the plasma membrane in bacteria requires a channel formed from the SecY complex, which cooperates with either a translating ribosome in cotranslational translocation or the SecA ATPase in post-translational translocation. Whether translocation requires oligomers of the SecY complex is an important but controversial issue: it determines channel size, how the permeation of small molecules is prevented, and how the channel interacts with the ribosome and SecA. Here, we probe in vivo the oligomeric state of SecY by cross-linking, using defined co- and post-translational translocation intermediates in intact Escherichia coli cells. We show that nontranslocating SecY associated transiently through different interaction surfaces with other SecY molecules inside the membrane. These interactions were significantly reduced when a translocating polypeptide inserted into the SecY channel co- or post-translationally. Mutations that abolish the interaction between SecY molecules still supported viability of E. coli. These results show that a single SecY molecule is sufficient for protein translocation.
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