Cotranslational folding inhibits translocation from within the ribosome-Sec61 translocon complex

Brian J. Conti; Johannes Elferich; Zhongying Yang; Ujwal Shinde; William R. Skach

doi:10.1038/nsmb.2779

Cotranslational folding inhibits translocation from within the ribosome-Sec61 translocon complex

Brian J. Conti
, Johannes Elferich
, Zhongying Yang
, Ujwal Shinde
, William R. Skach

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

Eukaryotic secretory proteins cross the endoplasmic reticulum (ER) membrane through a protein-conducting channel contained within the ribosome- Sec61translocon complex (RTC). Using a zinc-finger sequence as a folding switch, we show that cotranslational folding of a secretory passenger inhibits translocation in canine ER microsomes and in human cells. Folding occurs within a cytosolically inaccessible environment, after ER targeting but before initiation of translocation, and it is most effective when the folded domain is 15-54 residues beyond the signal sequence. Under these conditions, substrate is diverted into cytosol at the stage of synthesis in which unfolded substrate enters the ER lumen. Moreover, the translocation block is reversed by passenger unfolding even after cytosol emergence. These studies identify an enclosed compartment within the assembled RTC that allows a short span of nascent chain to reversibly abort translocation in a substrate-specific manner.

Original language	English (US)
Pages (from-to)	228-235
Number of pages	8
Journal	Nature Structural and Molecular Biology
Volume	21
Issue number	3
DOIs	https://doi.org/10.1038/nsmb.2779
State	Published - Jan 1 2014

Funding

We thank V. Hilser, P. Devaraneni and members of the Skach laboratory for valuable discussion. This work was supported by US National Institutes of Health grants GM53457 (W.R.S.), DK51818 (W.R.S.), F32 GM083568 (B.J.C.) and T32 HL083808 (B.J.C.), by the Cystic Fibrosis Foundation Therapeutics (W.R.S.) and by US National Science Foundation grant MCB0746589 (U.S.) and American Heart Association grant 12PRE11470005 (J.E.).

Funders	Funder number
Author National Science Foundation National Science Foundation National Institutes of Health National Institutes of Health National Institutes of Health National Institutes of Health National Science Foundation National Science Foundation	MCB0746589
Author National Institutes of Health National Institutes of Health National Institutes of Health National Institutes of Health The Bev Hartig Huntington's Disease Foundation National Institutes of Health	GM53457, DK51818, F32 GM083568
National Institute of Health National Heart, Lung, and Blood Institute	T32HL083808
American Heart Association/American Stroke Association	12PRE11470005
Cystic Fibrosis Foundation Therapeutics

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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10.1038/nsmb.2779

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title = "Cotranslational folding inhibits translocation from within the ribosome-Sec61 translocon complex",

abstract = "Eukaryotic secretory proteins cross the endoplasmic reticulum (ER) membrane through a protein-conducting channel contained within the ribosome- Sec61translocon complex (RTC). Using a zinc-finger sequence as a folding switch, we show that cotranslational folding of a secretory passenger inhibits translocation in canine ER microsomes and in human cells. Folding occurs within a cytosolically inaccessible environment, after ER targeting but before initiation of translocation, and it is most effective when the folded domain is 15-54 residues beyond the signal sequence. Under these conditions, substrate is diverted into cytosol at the stage of synthesis in which unfolded substrate enters the ER lumen. Moreover, the translocation block is reversed by passenger unfolding even after cytosol emergence. These studies identify an enclosed compartment within the assembled RTC that allows a short span of nascent chain to reversibly abort translocation in a substrate-specific manner.",

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Cotranslational folding inhibits translocation from within the ribosome-Sec61 translocon complex

Abstract

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