2G3K

Crystal structure of the C-terminal domain of Vps28

Summary for 2G3K

• Entry DOI: 10.2210/pdb2g3k/pdb
• Descriptor: Vacuolar protein sorting-associated protein VPS28 (2 entities in total)
• Functional Keywords: 4 helix bundle, transport protein
• Biological source: Saccharomyces cerevisiae (baker's yeast)
• Cellular location: Cytoplasm: Q02767
• Total number of polymer chains: 7
• Total formula weight: 76667.03

Authors

Pineda-Molina, E.,Belrhali, H.,Piefer, A.J.,Akula, I.,Bates, P.,Weissenhorn, W. (deposition date: 2006-02-20, release date: 2006-06-27, Last modification date: 2024-10-30)

Primary citation

Pineda-Molina, E.,Belrhali, H.,Piefer, A.J.,Akula, I.,Bates, P.,Weissenhorn, W.
The crystal structure of the C-terminal domain of Vps28 reveals a conserved surface required for Vps20 recruitment.
Traffic, 7:1007-1016, 2006

PubMed Abstract: The endosomal sorting complex I required for transport (ESCRT-I) is composed of the three subunits Vps23/Tsg101, Vps28 and Vps37. ESCRT-I is recruited to cellular membranes during multivesicular endosome biogenesis and by enveloped viruses such as HIV-1 to mediate budding from the cell. Here, we describe the crystal structure of a conserved C-terminal domain from Sacharomyces cerevisiae Vps28 (Vps28-CTD) at 3.05 A resolution which folds independently into a four-helical bundle structure. Co-expression experiments of Vps28-CTD, Vps23 and Vps37 suggest that Vps28-CTD does not directly participate in ESCRT-I assembly and may thus act as an adaptor module for downstream interaction partners. We show through mutagenesis studies that Vps28-CTD employs its strictly conserved surface in the interaction with the ESCRT-III factor Vps20. Furthermore, we present evidence that Vps28-CTD is sufficient to rescue an equine infectious anaemia virus (EIAV) Gag late domain deletion. Vps28-CTD mutations abolishing Vps20 interaction in vitro also prevent the rescue of the EIAV Gag late domain mutant consistent with a potential direct Vps28-ESCRT-III Vps20 recruitment. Therefore, the physiological relevant EIAV Gag-Alix interaction can be functionally replaced by a Gag-Vps28-CTD fusion. Because both Alix and Vps28-CTD can directly recruit ESCRT-III proteins, ESCRT-III assembly coupled to Vps4 action may therefore constitute the minimal budding machinery for EIAV release.

PubMed: 16749904
DOI: 10.1111/j.1600-0854.2006.00440.x

Experimental method

X-RAY DIFFRACTION (3.05 Å)