NMR Structure of the N-Terminal Domain of Capsid Protein from the Mason-Pfizer Monkey Virus |
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Authors: | Pavel Macek,Josef Chmelí k,Ivana K?í ?ová ,Pavel Kade?á vek,Luká &scaron ?í dek,Romana Hadravová ,Iva Pichová ,Michaela Rumlová ,Vladimí r Sklená ? |
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Affiliation: | 1 National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kotlá?ská 2, 611 37 Brno, Czech Republic 2 Gilead Sciences and IOCB Research Centre, Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague, Czech Republic 3 Loschmidt Laboratories, Institute of Experimental Biology and National Centre for Biomolecular Research, Masaryk University, Kotlá?ská 2, 611 37 Brno, Czech Republic 4 Institute of Chemical Technology, Technická 5, 166 28 Prague, Czech Republic 5 Institute of Microbiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic |
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Abstract: | The high-resolution structure of the N-terminal domain (NTD) of the retroviral capsid protein (CA) of Mason-Pfizer monkey virus (M-PMV), a member of the betaretrovirus family, has been determined by NMR. The M-PMV NTD CA structure is similar to the other retroviral capsid structures and is characterized by a six α-helix bundle and an N-terminal β-hairpin, stabilized by an interaction of highly conserved residues, Pro1 and Asp57. Since the role of the β-hairpin has been shown to be critical for formation of infectious viral core, we also investigated the functional role of M-PMV β-hairpin in two mutants (i.e., ΔP1NTDCA and D57ANTDCA) where the salt bridge stabilizing the wild-type structure was disrupted. NMR data obtained for these mutants were compared with those obtained for the wild type. The main structural changes were observed within the β-hairpin structure; within helices 2, 3, and 5; and in the loop connecting helices 2 and 3. This observation is supported by biochemical data showing different cleavage patterns of the wild-type and the mutated capsid-nucleocapsid fusion protein (CANC) by M-PMV protease. Despite these structural changes, the mutants with disrupted salt bridge are still able to assemble into immature, spherical particles. This confirms that the mutual interaction and topology within the β-hairpin and helix 3 might correlate with the changes in interaction between immature and mature lattices. |
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Keywords: | M-PMV, Mason-Pfizer monkey virus HIV-1, human immunodeficiency virus 1 NTD, N-terminal domain RSV, Rous sarcoma virus HTLV, human T-cell leukemia virus EIAV, equine infectious anemia virus B-MLV, B-tropic murine leukemia virus JSRV, Jaagsiekte sheep retrovirus WT, wild-type NOE, nuclear Overhauser enhancement RDCs, residual dipolar couplings ssNOE, steady-state {1H}-15N NOE enhancement CCSP, combined chemical shift perturbations |
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