کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2188349 | 1096163 | 2007 | 13 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Mutations in Capsid Major Homology Region Affect Assembly and Membrane Affinity of HIV-1 Gag Mutations in Capsid Major Homology Region Affect Assembly and Membrane Affinity of HIV-1 Gag](/preview/png/2188349.png)
We introduced mutations into the HIV-1 major homology region (MHR; capsids 153–172) and adjacent C-terminal region to analyze their effects on virus-like particle (VLP) assembly, membrane affinity, and the multimerization of the Gag structural protein. Results indicate that alanine substitutions at K158, F168 or E175 significantly diminished VLP production. All assembly-defective Gag mutants had markedly reduced membrane-binding capacities, but results from a velocity sedimentation analysis suggest that most of the membrane-bound Gag proteins were present, primarily in a higher-order multimerized form. The membrane-binding capacity of the K158A, F168A, and E175A Gag proteins increased sharply upon removal of the MA globular domain. While demonstrating improved multimerization capability, the two MA-deleted versions of F168A and E175A did not show marked improvement in VLP production, presumably due to a defect in association with the raft-like membrane domain. However, K158A bound to detergent-resistant raft-like membrane; this was accompanied by noticeably improved VLP production following MA removal. Our results suggest that the HIV-1 MHR and adjacent downstream region facilitate multimerization and tight Gag packing. Enhanced Gag multimerization may help expose the membrane-binding domain and thus improve Gag membrane binding, thereby promoting Gag multimerization into higher-order assembly products.
Journal: Journal of Molecular Biology - Volume 370, Issue 3, 13 July 2007, Pages 585–597