HIV Gag mimics the Tsg101-recruiting activity of the human Hrs protein.
The HIV-1 Gag protein recruits the cellular factor Tsg101 to facilitate the final stages of virus budding. A conserved P(S/T)AP tetrapeptide motif within Gag (the "late domain") binds directly to the NH2-terminal ubiquitin E2 variant (UEV) domain of Tsg101. In the cell, Tsg101 is required for biogenesis of vesicles that bud into the lumen of late endosomal compartments called multivesicular bodies (MVBs). However, the mechanism by which Tsg101 is recruited from the cytoplasm onto the endosomal membrane has not been known. Now, we report that Tsg101 binds the COOH-terminal region of the endosomal protein hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs; residues 222-777). This interaction is mediated, in part, by binding of the Tsg101 UEV domain to the Hrs 348PSAP351 motif. Importantly, Hrs222-777 can recruit Tsg101 and rescue the budding of virus-like Gag particles that are missing native late domains. These observations indicate that Hrs normally functions to recruit Tsg101 to the endosomal membrane. HIV-1 Gag apparently mimics this Hrs activity, and thereby usurps Tsg101 and other components of the MVB vesicle fission machinery to facilitate viral budding.
Pornillos
Owen
O
Department of Biochemistry, University of Utah, School of Medicine, Salt Lake City, UT 84132, USA. wes@biochem.utah.edu
Higginson
Daniel S
DS
Stray
Kirsten M
KM
Fisher
Robert D
RD
Garrus
Jennifer E
JE
Payne
Marielle
M
He
Gong-Ping
GP
Wang
Hubert E
HE
Morham
Scott G
SG
Sundquist
Wesley I
WI
D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism*; D056827:Endosomal Sorting Complexes Required for Transport; D011992:Endosomes / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D005057:Eukaryotic Cells / Q000378:metabolism / Q000821:virology*; D015683:Gene Products, gag / Q000378:metabolism*; D006678:HIV / Q000378:metabolism* / Q000472:pathogenicity / Q000648:ultrastructure; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism / Q000648:ultrastructure; D008854:Microscopy, Electron; D018716:Molecular Mimicry / Q000502:physiology; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D014157:Transcription Factors / Q000378:metabolism*; D022161:Transport Vesicles / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology; D014779:Virus Replication / Q000502:physiology*; D017201:Virus Shedding / Q000502:physiology*; D054301:gag Gene Products, Human Immunodeficiency Virus; D000099177:Tsg101 Protein
D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D015683:Gene Products, gag; D010750:Phosphoproteins; D014157:Transcription Factors; D054301:gag Gene Products, Human Immunodeficiency Virus; D000099177:Tsg101 Protein; C083868:gag protein p1, Human immunodeficiency virus; C096164:hepatocyte growth factor-regulated tyrosine kinase substrate