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PMID: 23201121 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

ALIX is a Lys63-specific polyubiquitin binding protein that functions in retrovirus budding.

Developmental cell ·Vol. 23 ·No. 6 ·2012-12-11 ·Pages 1247-54

Dowlatshahi DP, Sandrin V, Vivona S, Shaler TA, Kaiser SE, Melandri F, Sundquist WI, Kopito RR

Abstract

The diversity of ubiquitin (Ub)-dependent signaling is attributed to the ability of this small protein to form different types of covalently linked polyUb chains and to the existence of Ub binding proteins that interpret this molecular syntax. We used affinity capture/mass spectrometry to identify ALIX, a component of the ESCRT pathway, as a Ub binding protein. We report that the V domain of ALIX binds directly and selectively to K63-linked polyUb chains, exhibiting a strong preference for chains composed of more than three Ub. Sequence analysis identified two potential Ub binding sites on a single α-helical surface within the coiled-coil region of the V domain. Mutation of these putative Ub binding sites inhibited polyUb binding to the isolated V domain in vitro and impaired budding of lentiviruses. These data reveal an important role for K63 polyUb binding by ALIX in retroviral release.

MeSH Terms
Binding Sites/genetics Calcium-Binding Proteins/chemistry,metabolism Carrier Proteins/chemistry,metabolism Cell Cycle Proteins/chemistry,metabolism Cell Line Endosomal Sorting Complexes Required for Transport/chemistry,metabolism HEK293 Cells HIV-1/physiology Humans Infectious Anemia Virus, Equine/physiology Protein Binding Protein Interaction Domains and Motifs Protein Structure, Secondary Retroviridae/physiology Signal Transduction Ubiquitin/chemistry,metabolism Virus Release
Chemicals
Calcium-Binding Proteins Carrier Proteins Cell Cycle Proteins Endosomal Sorting Complexes Required for Transport PDCD6IP protein, human Ubiquitin
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Dowlatshahi Dara P
Department of Biology, Stanford University, Stanford, CA 94305, USA.
Sandrin Virginie
Vivona Sandro
Shaler Thomas A
Kaiser Stephen E
Melandri Francesco
Sundquist Wesley I
Kopito Ron R
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Article Info
Journal
Developmental cell
Abbr.
Dev Cell
ISSN
1878-1551
DOI
10.1016/j.devcel.2012.10.023
Published
2012-12-11
Epub
2012-00-29
Pages
1247-54
Language
English
Region
United States
NLM ID
101120028
PMCID
PMC3522770
Subset
IM
Grants
NIAID NIH HHS · R01 AI051174 · United States
NIGMS NIH HHS · R01 GM074874 · United States
External Links
PubMed source DOI: 10.1016/j.devcel.2012.10.023 PMC Full Text View journal details
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