1H, 13C and 15N backbone and partial side-chain resonance assignments of the C-terminal domain of HIV-1 Pr55Gag encompassed in NCp15
- 83 Downloads
During HIV-1 assembly, the Pr55Gag polyprotein precursor (Gag) interacts with the genomic RNA, with lipids of the plasma membrane, with host proteins (ALIX, TSG101) through the ESCRT complex, with the viral protein Vpr and are involved in intermolecular interactions with other Pr55Gag proteins. This network of interactions is responsible for the formation of the viral particle, the selection of genomic RNA and the packaging of Vpr. The C-terminal domain of Gag encompassed in NCp15 is involved in the majority of these interactions, either by its nucleocapsid or its p6 domains. We study the NCp15 protein as a model of the C-terminal domain of Gag to better understand the role of this domain in the assembly and budding of HIV-1. Here, we report the 1H, 13C and 15N chemical shift assignments of NCp15 obtained by heteronuclear multidimensional NMR spectroscopy as well as the analysis of its secondary structure in solution. These assignments of NCp15 pave the way for interaction studies with its numerous partners.
KeywordsNucleocapsid HIV-1 NCp15 NMR Resonance assignment
This work was supported by grants from SIDACTION and ANRS. We thank Nelly Morellet and Ewen Lescop for technical advice and helpful discussion. Financial support from the TGIR-RMN-THC FR3050 CNRS for conducting the research is gratefully acknowledged. We are grateful to Feder, Sesame Ile-de-France and Paris Descartes University that financed a new NMR console that allows us to perform NMR experiments with state-of-the-art 600 MHz spectrometer.
- De Guzman RN, Wu ZR, Stalling CC, Pappalardo L, Borer PN, Summers MF (1998) Structure of the HIV-1 nucleocapsid protein bound to the SL3 psi-RNA. Recogn Elem Sci 279:384–388Google Scholar
- Dussupt V, Javid MP, Abou-Jaoude G, Jadwin JA, de La Cruz J, Nagashima K, Bouamr F (2009) The nucleocapsid region of HIV-1 Gag cooperates with the PTAP and LYPXnL late domains to recruit the cellular machinery necessary for viral budding. PLoS Pathog 5:e1000339. https://doi.org/10.1371/journal.ppat.1000339 CrossRefGoogle Scholar
- Fossen T, Wray V, Bruns K, Rachmat J, Henklein P, Tessmer U, Maczurek A, Klinger P, Schubert U (2005) Solution structure of the human immunodeficiency virus type 1 p6 protein. J Biol Chem 280:42515–42527. https://doi.org/10.1074/jbc.M507375200
- Goddard TD, Kneller DG SPARKY 3. University of Califormia, San FranciscoGoogle Scholar
- Morellet N, Jullian N, De Rocquigny H, Maigret B, Darlix JL, Roques BP (1992) Determination of the structure of the nucleocapsid protein NCp7 from the human immunodeficiency virus type 1 by 1H NMR. Embo J 11:3059–3065Google Scholar
- Salgado GF, Vogel A, Marquant R, Feller SE, Bouaziz S, Alves ID (2009b) The role of membranes in the organization of HIV-1 Gag p6 and Vpr: p6 shows high affinity for membrane bilayers which substantially increases the interaction between p6 and Vpr. J Med Chem 52:7157–7162. https://doi.org/10.1021/jm901106t CrossRefGoogle Scholar
- Solbak SM, Reksten TR, Hahn F, Wray V, Henklein P, Henklein P, Halskau Ø, Schubert U, Fossen T (2013) HIV-1 p6 - a structured to flexible multifunctional membrane-interacting protein. Biochim Biophys Acta 1828:816–823. https://doi.org/10.1016/j.bbamem.2012.11.010
- Vranken WF, Boucher W, Stevens TJ, Fogh RH, Pajon A, Llinas M, Ulrich EL, Markley JL, Ionides J, Laue ED (2005) The CCPN data model for NMR spectroscopy: development of a software pipeline. Proteins 59:687–696. https://doi.org/10.1002/prot.20449
- Wishart DS, Sykes BD, Richards FM (1992) The chemical shift index: a fast and simple method for the assignment of protein secondary structure through. NMR Spectrosc Biochem 31:1647–1651Google Scholar