The Mason-Pfizer monkey virus PPPY and PSAP motifs both contribute to virus release.
Late (L) domains are required for the efficient release of several groups of enveloped viruses. Three amino acid motifs have been shown to provide L-domain function, namely, PPXY, PT/SAP, or YPDL. The retrovirus Mason-Pfizer monkey virus (MPMV) carries closely spaced PPPY and PSAP motifs. Mutation of the PPPY motif results in a complete loss of virus release. Here, we show that the PSAP motif acts as an additional L domain and promotes the efficient release of MPMV but requires an intact PPPY motif to perform its function. Examination of HeLaP4 cells expressing PSAP mutant virus by electron microscopy revealed mostly late budding structures and chains of viruses accumulating at the cell surface with little free virus. In the case of the PPPY mutant virus, budding appeared to be mostly arrested at an earlier stage before induction of membrane curvature. The cellular protein TSG101, which interacts with the human immunodeficiency virus type 1 (HIV-1) PTAP L domain, was packaged into MPMV in a PSAP-dependent manner. Since TSG101 is crucial for HIV-1 release, this result suggests that the Gag-TSG101 interaction is responsible for the virus release function of the MPMV PSAP motif. Nedd4, which has been shown to interact with viral PPPY motifs, was also detected in MPMV particles, albeit at much lower levels. Consistent with a role of VPS4A in the budding of both PPPY and PTAP motif-containing viruses, the overexpression of ATPase-defective GFP-VPS4A fusion proteins blocked both wild-type and PSAP mutant virus release.
Gottwein
Eva
E
Abteilung Virologie, Universitätsklinikum Heidelberg, D-69120 Heidelberg, Germany.
Bodem
Jochen
J
Müller
Barbara
B
Schmechel
Ariane
A
Zentgraf
Hanswalter
H
Kräusslich
Hans-Georg
HG
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport; D015683:Gene Products, gag / Q000737:chemistry / Q000235:genetics / Q000502:physiology; D015497:HIV-1 / Q000235:genetics / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008025:Ligases / Q000378:metabolism; D016093:Mason-Pfizer monkey virus / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D000075702:Nedd4 Ubiquitin Protein Ligases; D011499:Protein Processing, Post-Translational; D017384:Sequence Deletion; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection; D044767:Ubiquitin-Protein Ligases; D019065:Virus Assembly; D000099177:Tsg101 Protein
D002135:Calcium-Binding Proteins; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D015683:Gene Products, gag; D008025:Ligases; D000075702:Nedd4 Ubiquitin Protein Ligases; D014157:Transcription Factors; D044767:Ubiquitin-Protein Ligases; D000099177:Tsg101 Protein; C000618483:Nedd4 protein, human