title: Structural basis for ESCRT-III CHMP3 recruitment of AMSH.
abstract: Endosomal sorting complexes required for transport (ESCRT) recognize ubiquitinated cargo and catalyze diverse budding processes including multivesicular body biogenesis, enveloped virus egress, and cytokinesis. We present the crystal structure of an N-terminal fragment of the deubiquitinating enzyme AMSH (AMSHΔC) in complex with the C-terminal region of ESCRT-III CHMP3 (CHMP3ΔN). AMSHΔC folds into an elongated 90 Å long helical assembly that includes an unusual MIT domain. CHMP3ΔN is unstructured in solution and helical in complex with AMSHΔC, revealing a novel MIT domain interacting motif (MIM) that does not overlap with the CHMP1-AMSH binding site. ITC and SPR measurements demonstrate an unusual high-affinity MIM-MIT interaction. Structural analysis suggests a regulatory role for the N-terminal helical segment of AMSHΔC and its destabilization leads to a loss of function during HIV-1 budding. Our results indicate a tight coupling of ESCRT-III CHMP3 and AMSH functions and provide insight into the regulation of ESCRT-III.
authors.affiliation: Unit of Virus Host Cell Interactions (UVHCI) UMI 3265 Université Joseph Fourier-EMBL-CNRS, 6 rue Jules Horowitz 38042 Grenoble Cedex 9, France.
authors.forename: Julianna
authors.initials: J
authors.lastname: Solomons
authors.forename: Charles
authors.initials: C
authors.lastname: Sabin
authors.forename: Emilie
authors.initials: E
authors.lastname: Poudevigne
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Usami
authors.forename: David Lutje
authors.initials: DL
authors.lastname: Hulsik
authors.forename: Pauline
authors.initials: P
authors.lastname: Macheboeuf
authors.forename: Bettina
authors.initials: B
authors.lastname: Hartlieb
authors.forename: Heinrich
authors.initials: H
authors.lastname: Göttlinger
authors.forename: Winfried
authors.initials: W
authors.lastname: Weissenhorn
mesh_terms: D000595:Amino Acid Sequence; D056827:Endosomal Sorting Complexes Required for Transport / Q000737:chemistry* / Q000378:metabolism; D057809:HEK293 Cells; D015658:HIV Infections / Q000821:virology; D015497:HIV-1 / Q000502:physiology; D006801:Humans; D006860:Hydrogen Bonding; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry* / Q000378:metabolism; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D043222:Ubiquitin Thiolesterase / Q000737:chemistry* / Q000378:metabolism; D057074:Virus Release
chemical_list: C498543:CHMP3 protein, human; D056827:Endosomal Sorting Complexes Required for Transport; D046912:Multiprotein Complexes; D010446:Peptide Fragments; C486696:STAMBP protein, human; D043222:Ubiquitin Thiolesterase