The NC domain of HIV-1 Gag contributes to the interaction of Gag with TSG101

Meshri, Salah Edin El; Boutant, Emmanuel; Mouhand, Assia; Thomas, Audrey; Larue, Valéry; Richert, Ludovic; Vivet-Boudou, Valérie; Tisné, Carine; Muriaux, Delphine; Rocquigny, Hugues de

doi:10.1016/j.bbagen.2018.03.020

Cited by 16 publications

(22 citation statements)

References 51 publications

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“…Using lysates from cells expressing non-coding HIV-1 gRNA, GST-tagged NCp15, GST-tagged NCp9, GST-tagged NCp15∆ZnF 2 or GST alone, Chamontin et al [17] identified ZnF 2 as the element in the protein critical for Tsg101 recognition. El Meshri et al identified several residues in NC that were perturbed when Tsg101 bound the PTAP motif in Gag, suggesting that NC contributes to the Gag-Tsg101 interaction [15].The study reported herein provides direct evidence that Tsg101 can bind RNA through its UEV domain and demonstrates that the interaction controls Tsg101 recruitment. RNA-binding might facilitate Gag recruitment by reducing its interaction with endosomal membranes in a manner analogous to how tRNA interaction with MA is believed to minimize its sequestering on internal membranes [41,54].…”

Section: Discussionmentioning

confidence: 54%

“…A model suggesting that RNA binding interferes with recognition of UEV Ub-binding sites important for productive Gag-Tsg101 interaction is proposed in Figure 9. motif in Gag, suggesting that NC contributes to the Gag-Tsg101 interaction [15].The study reported herein provides direct evidence that Tsg101 can bind RNA through its UEV domain and demonstrates that the interaction controls Tsg101 recruitment. RNA-binding might facilitate Gag recruitment by reducing its interaction with endosomal membranes in a manner analogous to how tRNA interaction with MA is believed to minimize its sequestering on internal membranes [41,54].…”

Section: Discussionmentioning

confidence: 54%

“…Employing a chimeric protein to deliver Tsg101 to the assembly site circumvents technical limitations associated with "hyper"-expression of the Tsg101 protein arising from the tight cellular control of its steady-state level [47]. Their findings and the subsequent studies conducted by El Meshri et al [15] showed that Tsg101 recruitment requires contributions from both the NC and p6 domains. We confirmed the finding that providing Tsg101 at the budding site by translational fusion of Tsg101 to Gag-∆ZnF 2 (creating Gag-∆ZnF 2 In all cases, the cell lysate signal was normalized for total protein content using the cellular protein actin as an internal marker (panel B, bottom).…”

Section: The Uev Domain Is Not Required When Tsg101 Is At the Buddingmentioning

confidence: 99%

“…Tsg101 binds only p6 [11][12][13]. However, mutations of basic residues throughout NC or in zinc fingers (ZnFs) cause a significant decline in release of particles carrying the intact PTAP motif indicating that, in the context of NC mutants, the PTAP motif is not sufficient to drive virus release [8,9,14,15]. These findings collectively indicated that NC and the PTAP motif play cooperative roles in viral egress.…”

Section: Introductionmentioning

confidence: 99%

“…This group further demonstrated that Tsg101, when substituted for ZnF2, rescued the budding defect caused by ZnF2 deletion. By employing a combination of the fluorescence resonance energy transfer (FLIM-FRET) technique and NMR, El Meshri et al subsequently showed that deletion of the entire NC domain, or even just the two zinc finger motifs within this domain, alters the Gag-directed cellular distribution of Tsg101 and negatively impacts their interaction [15].…”

Section: Introductionmentioning

confidence: 99%

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RNA Binding Suppresses Tsg101 Recognition of Ub-Modified Gag and Facilitates Recruitment to the Plasma Membrane

Watanabe

Strickland

Tjandra

et al. 2020

Viruses

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The ESCRT-I factor Tsg101 is essential for sorting endocytic cargo and is exploited by viral pathogens to facilitate egress from cells. Both the nucleocapsid (NC) domain and p6 domain in HIV-1 Gag contribute to recruitment of the protein. However, the role of NC is unclear when the P(S/T)AP motif in p6 is intact, as the motif recruits Tsg101 directly. The zinc fingers in NC bind RNA and membrane and are critical for budding. Tsg101 can substitute for the distal ZnF (ZnF2) and rescue budding of a mutant made defective by deletion of this element. Here, we report that the ubiquitin (Ub) E2 variant (UEV) domain in Tsg101 binds tRNA in vitro. We confirmed that Tsg101 can substitute for ZnF2 when provided at the viral assembly site as a chimeric Gag-Tsg101 protein (Gag-∆ZnF2-Tsg101) and rescue budding. The UEV was not required in this context; however, mutation of the RNA binding determinants in UEV prevented Tsg101 recruitment from the cell interior when Gag and Tsg101 were co-expressed. The same Tsg101 mutations increased recognition of Gag-Ub, suggesting that tRNA and Ub compete for binding sites. This study identifies a novel Tsg101 binding partner that may contribute to its function in recognition of Ub-modified cargo.

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Section: Discussionmentioning

confidence: 54%

Section: Discussionmentioning

confidence: 54%

Section: The Uev Domain Is Not Required When Tsg101 Is At the Buddingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

RNA Binding Suppresses Tsg101 Recognition of Ub-Modified Gag and Facilitates Recruitment to the Plasma Membrane

Watanabe

Strickland

Tjandra

et al. 2020

Viruses

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Lysosomes Targeting pH Activable Imaging‐Guided Photodynamic Agents

et al. 2023

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Photodynamic therapy (PDT) is a photochemistry-based medical treatment combining light at a specific wavelength and a photosensitizer (PS) in the presence of oxygen. Application of PDT as a conventional treatment is limited and clearly the approval in clinics of new PS is challenging. The selective accumulation of the PS in the targeted malignant cells is of paramount importance to reduce the side effects that are typical of the current worldwide approved PS. Here we report a new series of aniline-and iodine-substituted BODIPY derivatives (1-3) as promising lysosome-targeting and pH-responsive theranostic PS, which displayed a significant in vitro lightinduced cytotoxicity, efficient imaging properties and low dark toxicity (for 2 and 3). These compounds were obtained in few reproducible synthetic steps and good yields. Spectroscopic and electrochemical measurements along with computational calculations confirmed the quenching of the emissive properties of the PS, while both fluorescence and 1 O 2 emission were obtained only under acidic conditions inducing amine protonation. The pK a values and pH-dependent emissive properties of 1-3 being established, their cellular uptake and activation in the lysosomal vesicles (pH � 4-5) were confirmed by their colocalization with the commercial LysoTracker deep red and light-induced cytotoxicity (IC 50 between 0.16 and 0.06 μM) against HeLa cancer cells.

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