Abstract
The X-ray crystallographic structure of HIV-1 capsid protein suggests that the dimer interface of the dimerization domain is mainly formed from a putative α-helix structure of 14 amino acids (Gag residues 311–324) and lies directly C-terminal to the capsid major homology region. We found that a deletion mutation in the α-helix drastically reduces virus particle production. Alanine-scanning mutagenetic analysis indicated that substitution mutations at residues Q311, V313, K314, W316, and M317 all impair virus particle production markedly. Membrane flotation assays suggested that some mutations in the dimer interface have slight effects on the efficient binding of Gag to membranes. Indirect immunofluorescence studies revealed that mutants defective in virus production exhibit a subcellular distribution pattern similar to that of wild-type. However, velocity sedimentation analysis showed that mutations significantly impairing virus particle production were also detrimental to Gag multimerization, suggesting that the impaired virus production may be due to a defect in Gag multimerization. These results support the proposal that residues in the capsid dimer interface play a crucial role in promoting Gag multimerization, possibly by facilitating stable Gag–Gag interactions.
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Acknowledgements
We thank H.-C. Chiu and W.-H. Liao for reagents and technical assistance. The hybridoma clone 183 H12-5C was provided by Bruce Chesebro of the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, MD. This work was supported by grants VGH93-309 from the Taipei Veterans General Hospital and NSC93-2320-B-010–015 from the National Science Council, Taiwan, Republic of China
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Chu, HH., Chang, YF. & Wang, CT. Mutations in the α-helix Directly C-terminal to the Major Homology Region of Human Immunodeficiency Virus Type 1 Capsid Protein Disrupt Gag Multimerization and Markedly Impair Virus Particle Production. J Biomed Sci 13, 645–656 (2006). https://doi.org/10.1007/s11373-006-9094-6
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DOI: https://doi.org/10.1007/s11373-006-9094-6