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Tsg101/ESCRT-I Recruitment Regulated by the Dual Binding Modes of K63-Linked Diubiquitin

40 Pages Posted: 20 Feb 2020 Publication Status: Review Complete

See all articles by Madeleine Strickland

Madeleine Strickland

National Institutes of Health - Biochemistry and Biophysics Center

Susan Watanabe

Stony Brook University - Department of Microbiology and Immunology

Steven M. Bonn

University of Maryland - Department of Chemistry and Biochemistry

Christina M. Camara

University of Maryland - Department of Chemistry and Biochemistry

David Fushman

University of Maryland - Department of Chemistry and Biochemistry

Carol A. Carter

State University of New York (SUNY) - Department of Microbiology and Immunology

Nico Tjandra

National Institutes of Health - Biochemistry and Biophysics Center

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Abstract

The ESCRT-I protein Tsg101 plays a critical role in viral budding and endocytic sorting. Although Tsg101 is known to recognize monoubiquitin (Ub1), here we show for the first time that it can also bind several diubiquitins (Ub2), with a preference for K63-linked Ub2. The NMR structure of the Tsg101:K63-Ub2 complex showed that while the Ub1-binding site accommodates the distal domain of Ub2, the proximal domain alternatively binds two different sites, the vestigial active site and an N-terminal helix. Mutation of each site resulted in a distinct phenotype with respect to ubiquitin-dependent recruitment. Mutation in the vestigial active site abrogated the interaction between Tsg101 and the HIV-1 protein Gag, while mutation at the N-terminal helix increased the population of Gag-Tsg101 in the cell interior. Given the broad involvement of Tsg101 in diverse cellular functions, this discovery advances our understanding of how the ESCRT protein recognizes binding partners and sorts endocytic cargo.

Keywords: ESCRT, Diubiquitin, Tsg101, NMR, CPMG, viral budding, endocytic sorting

Suggested Citation

Strickland, Madeleine and Watanabe, Susan and Bonn, Steven M. and Camara, Christina M. and Fushman, David and Carter, Carol A. and Tjandra, Nico, Tsg101/ESCRT-I Recruitment Regulated by the Dual Binding Modes of K63-Linked Diubiquitin. Available at SSRN: https://ssrn.com/abstract=3537944 or http://dx.doi.org/10.2139/ssrn.3537944
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Madeleine Strickland

National Institutes of Health - Biochemistry and Biophysics Center ( email )

Bethesda, MD 20892
United States

Susan Watanabe

Stony Brook University - Department of Microbiology and Immunology ( email )

United States

Steven M. Bonn

University of Maryland - Department of Chemistry and Biochemistry ( email )

United States

Christina M. Camara

University of Maryland - Department of Chemistry and Biochemistry ( email )

United States

David Fushman

University of Maryland - Department of Chemistry and Biochemistry ( email )

United States

Carol A. Carter

State University of New York (SUNY) - Department of Microbiology and Immunology ( email )

United States

Nico Tjandra (Contact Author)

National Institutes of Health - Biochemistry and Biophysics Center ( email )

Bethesda, MD 20892
United States

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