PPPYEPTAP Motif Is the Late Domain of Human T-Cell Leukemia Virus Type 1 Gag and Mediates Its Functional Interaction with Cellular Proteins Nedd4 and Tsg101

Bouamr, Fadila; Melillo, Jessica A.; Wang, Margaret Q.; Nagashima, Kunio; Santos, Martha de Los; Rein, Alan; Goff, Stephen P.

doi:10.1128/jvi.77.22.11882-11895.2003

Cited by 122 publications

(80 citation statements)

References 67 publications

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“…It has been recently suggested that Tsg101 and Nedd4 might respectively act at an early and late stage of HTLV-1 budding (Bouamr et al, 2003). Our data clearly support the opposite model as we obtained strong evidence that HTLV-1 Gag encounters Nedd4.1 before Tsg101.…”

Section: Discussionsupporting

confidence: 81%

“…By depleting Tsg101 in HTLV-1-producing cells, we demonstrated that this protein is also necessary for HTLV-1 release. Consistently, HTLV-1 Gag is able to associate with Tsg101 in vitro (Bouamr et al, 2003).…”

Section: Discussionsupporting

confidence: 57%

“…Our results are not consistent with this model as they reveal that the efficient release of HTLV-1 requires the integrity of both the PPPY and the PTAP motifs, as recently reported (Bouamr et al, 2003). MPMV, another oncoretrovirus, also uses both a PPPY and a PSAP motifs for budding (Gottwein et al, 2003).…”

Section: Discussioncontrasting

confidence: 56%

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Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 Gag trafficking towards the multivesicular body pathway prior to virus budding

Blot

Perugi

Prévost

et al. 2004

Journal of Cell Science

131

110

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One of the most exciting recent developments in the field of retroviruses is the finding that their Gag proteins hijack cellular proteins from the mutivesicular body (MVB) pathway during the budding process. The Gag proteins of oncoretroviruses possess a PPxY motif that recruits a ubiquitin ligase from the Nedd4 family, whereas those of the human immunodeficiency virus interact through a PTAP motif with Tsg101, a protein of the ESCRT-1 complex. It is currently assumed that Nedd4 and Tsg101 represent equivalent entry gates towards the same cellular process leading to budding, and that both partners are recruited to the plasma membrane where viral budding occurs. However, we report here that the budding of the human oncoretrovirus HTLV-1, the Gag proteins of which possess tandem PPPY/PTAP motifs, requires both Nedd4 and Tsg101. We show that Nedd4.1, but not Nedd4.2, is recruited by the PPPY motif of Gag and subsequently catalyzes Gag ubiquitination. We also demonstrate that Gag interacts first with Nedd4.1 at the plasma membrane and then with Tsg101 in late endosomes/MVBs. Consistently, we found that HTLV-1 particles mutated in the PPPY motif remain underneath the plasma membrane, blocked at an early step of the budding process, whereas PTAP-mutated viruses accumulate in intracellular vesicles, blocked at a later step. Our findings indicate that Nedd4.1 and Tsg101 act successively in the assembly process of HTLV-1 to ensure proper Gag trafficking through the endocytic pathway up to late endosomes where the late steps of retroviral release occur.

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Section: Discussionsupporting

confidence: 81%

Section: Discussionsupporting

confidence: 57%

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Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 Gag trafficking towards the multivesicular body pathway prior to virus budding

Blot

Perugi

Prévost

et al. 2004

Journal of Cell Science

131

110

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“…In this model, the viral protein Gag is the equivalent of the late endosomal cargo and different L-domains recruit the ESCRT proteins through the interaction with different cellular adaptor proteins. It is widely accepted that PTAP motifs recruit the ESCRT-I complex trough the interaction with TSG101 (28,35,36), whereas PPXY motifs utilize a subset of HECT ubiquitin ligases to facilitate viral budding (23,(37)(38)(39). A third type of L-domain containing the LYPXL motif recruits the ESCRT machinery by interacting with AIP1/ALIX (6, 7), the human homologue of Bro1.…”

Section: Interaction Of Amsh With the Class E Vps Pathway-basedmentioning

confidence: 99%

Interaction of AMSH with ESCRT-III and Deubiquitination of Endosomal Cargo

Agromayor

Martín-Serrano

2006

Journal of Biological Chemistry

116

117

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The "class E" vacuolar protein sorting (VPS) pathway mediates sorting of ubiquitinated cargo into the forming vesicles of the multivesicular bodies (MVB), and it is essential for down-regulation of signaling by growth factors and budding of enveloped viruses such as Ebola and HIV-1. Work in yeast has identified DOA4 as a gene that is recruited by the class E machinery to remove ubiquitin from the endosomal cargo before it is incorporated into MVB vesicles, but the identity of the mammalian counterpart is unclear. Here we report the interaction of AMSH (associated molecule with the SH3 domain of STAM), an endosomal deubiquitinating enzyme, with the endodomal sorting complex required for transport (ESCRT-III) subunits CHMP1A, CHMP1B, CHMP2A, and CHMP3. We also show that a catalytically inactive AMSH inhibits retroviral budding in a dominant-negative manner and induces the accumulation of ubiquitinated forms of an endosomal cargo, namely murine leukemia virus Gag. Finally, VPS4 and AMSH compete for binding to the C-terminal regions of CHMP1A and CHMP1B, revealing a coordinated interaction with ESCRT-III. Taken together, these results are consistent with a role of AMSH in the deubiquitination of the endosomal cargo preceding lysosomal degradation.Ubiquitin plays an essential role in the trafficking of membrane proteins into the degradative lysosomal pathway. Covalent attachment of ubiquitin to certain activated cell surface receptors serves as a sorting signal for entry into the endocytic vesicles at the plasma membrane (1). Subsequently to this internalization, the class E vacuolar protein sorting (VPS) 2 pathway directs the ubiquitinated cargo into the forming vesicles of the late endosomal compartment, termed multivesicular bodies (MVB) (2, 3). In the last step, the MVB fuses with the lysosomal membrane and the cargo is delivered to the lumen of the lysosome for degradation.Work in yeast has identified 18 class E VPS proteins that are required for sorting into the lumen of MVBs (2, 4, 5). Importantly, at least one human homolog for every class E protein has been identified and recent studies show that most of the protein-protein interactions between the class E proteins are conserved from yeasts to humans (6 -8). A subset of class E proteins form one of three endosomal sorting complexes required for transport (ESCRT-I, -II, and -III) (9 -11), and current models propose that these complexes are sequentially recruited by the endosomal cargo, forming a membrane-associated lattice that functions in vesicle invagination during MVB biogenesis. ESCRT-III is believed to contain the core sorting machinery of the class E pathway and it is composed of two functional subcomplexes, a membrane-proximal complex that interacts with the endosomal membrane and a peripheral subcomplex that seems to recruit accessory proteins (9).After completion of cargo sorting and vesicle formation, ESCRT-III recruits the AAA ATPase Vps4 through direct protein-protein interactions, mediating the disassembly of the ESCRT machinery for recycling in...

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“…Further analysis revealed that Nedd4 was expressed at various levels in several embryonic and adult tissues (42). Nedd4 is the prototypical member of the Nedd4 ubiquitin ligase family and regulates diverse cellular processes including signal transduction (which is involved in cancer development [10,49,73,75]), protein trafficking (52, 63), and viral budding (6,23,36,60,71,80).…”

mentioning

confidence: 99%

Herpes Simplex Virus Type 2 UL56 Interacts with the Ubiquitin Ligase Nedd4 and Increases Its Ubiquitination

Ushijima

Koshizuka

Goshima

et al. 2008

J Virol

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The herpes simplex virus UL56 gene is conserved among most members of the Alphaherpesvirinae family and plays a critical role in viral pathogenicity in vivo. The HSV-2 UL56 protein (UL56) is a C-terminally anchored type II membrane protein that is predicted to be inserted into the virion envelope, leaving its N-terminal domain in the tegument. UL56 interacts with KIF1A and UL11. Here we report that UL56 also interacts with the ubiquitin ligase Nedd4 and increases its ubiquitination. Nedd4 was identified as a UL56-interacting protein by a yeast two-hybrid screen. UL56 bound to Nedd4 via its PY motifs. Nedd4 was phosphorylated and degraded in wild-type HSV-2-infected cells but not in cells infected with a UL56-deficient mutant. Ubiquitination assays revealed that UL56 increased ubiquitinated Nedd4, which was actively degraded in infected cells. UL56 also caused a decrease in Nedd4 protein levels and the increased ubiquitination in cotransfected cells. However, UL56 itself was not ubiquitinated, despite its interaction with Nedd4. Based on these findings, we propose that UL56 regulates Nedd4 in HSV-2-infected cells, although deletion of UL56 had no apparent effect on viral growth in vitro.Herpes simplex virus (HSV) is a large, enveloped DNA virus with a genome possessing at least 74 different genes (19,46). Although approximately half of the HSV genes are not essential for replication in vitro, all of these accessory gene products are predicted to play indispensable roles in viral replication and dissemination in vivo (57).The HSV UL56 gene is an accessory gene that most members of the Alphaherpesvirinae family possess homologs for (except bovine herpes virus-1 and -5) (1,17,19,22,29,34,37,46,53,(65)(66)(67)(68)(69)(70); M. Schwyzer, V. Paces, G. J. Letchworth, V. Misra, H. J. Buhk, D. E. Lowery, C. Simard, L. J. Bello, E. Thiry, and C. Vlcek, 1995, complete DNA sequence of bovine herpesvirus 1. GenBank database [http://www.ncbi.nlm.nih.gov/Genbank/index .html] accession number NC_001847) and has been shown to play an important role in HSV type 1 (HSV-1) pathogenicity in vivo. UL56-deficient HSV-1 mutants are substantially less neuroinvasive (4, 58), although little is known molecularly about how this attenuation occurs. HSV-2 UL56 is a 235-amino-acid, C-terminally anchored, type II membrane protein (39) that is predicted to be inserted into the viral envelope so that the N-terminal domain is located in the virion tegument. In this topology, UL56 is predicted to have a 216-amino-acid cytoplasmic domain. UL56 associates with the neuron-specific kinesin KIF1A (41) and the HSV-2 protein UL11 (40). KIF1A is involved in the axonal transport of synaptic vesicle precursors (51), and its association with UL56 suggests that UL56 may affect vesicular transport in infected neurons. UL11 is a tegument protein that is involved in the envelopment and egress of viral nucleocapsids (3) and has dynamic membrane-trafficking properties (45). UL56 may specifically promote UL11 function in virion envelopment and egress in infected neur...

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PPPYEPTAP Motif Is the Late Domain of Human T-Cell Leukemia Virus Type 1 Gag and Mediates Its Functional Interaction with Cellular Proteins Nedd4 and Tsg101

Cited by 122 publications

References 67 publications

Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 Gag trafficking towards the multivesicular body pathway prior to virus budding

Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV-1 Gag trafficking towards the multivesicular body pathway prior to virus budding

Interaction of AMSH with ESCRT-III and Deubiquitination of Endosomal Cargo

Herpes Simplex Virus Type 2 UL56 Interacts with the Ubiquitin Ligase Nedd4 and Increases Its Ubiquitination

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