DOI: 10.1083/jcb.150.1.53. Role of the Cytoplasmic Segments of Sec61α in the Ribosome-Binding and Translocation-Promoting Activities of the Sec61 Complex

Role of the Cytoplasmic Segments of Sec61α in the Ribosome-Binding and Translocation-Promoting Activities of the Sec61 Complex

10.1083/jcb.150.1.53

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

The Sec61 complex performs a dual function in protein translocation across the RER, serving as both the high affinity ribosome receptor and the translocation channel. To define regions of the Sec61 complex that are involved in ribosome binding and translocation promotion, ribosome-stripped microsomes were subjected to limited digestions using proteases with different cleavage specificities. Protein immunoblot analysis using antibodies specific for the NH2 and COOH terminus of Sec61α was used to map the location of proteolysis cleavage sites. We observed a striking correlation between the loss of binding activity for nontranslating ribosomes and the digestion of the COOH- terminal tail or cytoplasmic loop 8 of Sec61α. The proteolyzed microsomes were assayed for SRP-independent translocation activity to determine whether high affinity binding of the ribosome to the Sec61 complex is a prerequisite for nascent chain transport. Microsomes that do not bind nontranslating ribosomes at physiological ionic strength remain active in SRP-independent translocation, indicating that the ribosome binding and translocation promotion activities of the Sec61 complex do not strictly correlate. Translocation-promoting activity was most severely inhibited by cleavage of cytosolic loop 6, indicating that this segment is a critical determinant for this function of the Sec61 complex.

Bibliography

Raden, D., Song, W., & Gilmore, R. (2000). Role of the Cytoplasmic Segments of Sec61Î± in the Ribosome-Binding and Translocation-Promoting Activities of the Sec61 Complex. The Journal of Cell Biology, 150(1), 53â64.

Authors 3

David Raden (first)
Weiqun Song (additional)
Reid Gilmore (additional)

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Dates

Type	When
Created	23 years ago (July 26, 2002, 12:46 p.m.)
Deposited	2 years, 1 month ago (July 22, 2023, 7:13 a.m.)
Indexed	1 month, 3 weeks ago (July 2, 2025, 2:31 p.m.)
Issued	25 years, 1 month ago (July 10, 2000)
Published	25 years, 1 month ago (July 10, 2000)
Published Online	25 years, 1 month ago (July 10, 2000)
Published Print	25 years, 1 month ago (July 10, 2000)

Funders 0

None

BibTeX

@article{Raden_2000, title={Role of the Cytoplasmic Segments of Sec61α in the Ribosome-Binding and Translocation-Promoting Activities of the Sec61 Complex}, volume={150}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.150.1.53}, DOI={10.1083/jcb.150.1.53}, number={1}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Raden, David and Song, Weiqun and Gilmore, Reid}, year={2000}, month=jul, pages={53–64} }

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