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Abstract
Crystal structure of the HIV-1 integrase catalytic core and C-terminal domains: a model for viral DNA binding.
Julian C.-H. Chen, Jolanta Krucinski, Larry J.W. Miercke, Janet S. Finer-Moore, Ann H. Tang, Andrew D. Leavitt, and Robert M. Stroud
Proc. of Natl. Acad. Sci. 97, 15 8233-8238, (2000).
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Insolubility of full-length HIV-1 integrase limited previous structure analyses to individual domains. By introducing five point mutations, we generated a more soluble integrase that allowed us to generate multi-domain HIV-1 integrase crystals. The first multi-domain HIV-1 integrase is reported. It incorporates the two-domain structure of the catalytic core and C-terminal domains (residues 52-288). The structure resolved to 2.8 is a Y-shaped dimer. Within the dimer, the core domains form the only dimer interface, and the C-terminal domain SH3-like folds are located 55 ? apart. A 26 amino acid a-helix, a6, links the C-terminal domain to the core domain. A kink in one of the two a6 helices occurs near a known proteolytic site suggesting that it may act as a flexible elbow to reorient the domains during the integration process. Other SH3-related b-sheets that contact DNA are structurally conserved in the HIV-1 integrase C-terminal domain suggesting a similar protein-DNA interaction in which the integrase C-terminal domain may serve to bind, bend, and orient viral DNA during integration. A strip of positively charged amino acids contributed by both monomers emerges from each active site of the dimer, suggesting a dimeric binding platform for binding the viral DNA ends. The crystal structure of the isolated catalytic core domain (residues 52-210), independently determined at 1.6 resolution, is identical to the core domain within the two-domain 52-288 structure.
Structure of HIV-1 IN52-288 (A) Stereoscopic view of the HIV-1 IN52-288 dimer, composed of monomer A (blue) and monomer B (green), demonstrate the monomer B active site in front. Monomer B catalytic residues D64, D116, and E152 are indicated (red balls), and the C- and N- termini of each monomer are labeled. The immunologically critical residue W235 is located on the surface. (B) Alignment of residues 195-270 in the dimer demonstrates the kink at T210 that creates a ~90o rotation of the C-terminal domains relative to one another as illustrated by P233 (blue dot). Figure generated by MOLSCRIPT [Kraulis, 1991 #408] and Raster3D [Merritt, 1994 #409]. |
