THE ANNEALING OF TRNA(3)(LYS) TO HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PRIMER BINDING-SITE IS CRITICALLY DEPENDENT ON THE NCP7 ZINC FINGERS STRUCTURE

Citation
E. Remy et al., THE ANNEALING OF TRNA(3)(LYS) TO HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 PRIMER BINDING-SITE IS CRITICALLY DEPENDENT ON THE NCP7 ZINC FINGERS STRUCTURE, The Journal of biological chemistry, 273(9), 1998, pp. 4819-4822
Citations number
36
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biology
ISSN journal
0021-9258
Volume
273
Issue
9
Year of publication
1998
Pages
4819 - 4822
Database
ISI
SICI code
0021-9258(1998)273:9<4819:TAOTTH>2.0.ZU;2-J
Abstract
The nucleocapsid protein NCp7 of the human immunodeficiency virus type 1 contains two zinc fingers of the CX2CX4HX4C type, flanked by severa l basic residues, and plays a major role in viral infectivity. Thus, N Cp7 was shown to promote annealing of the tRNA(3)(Lys) to the primer b inding site, a key step in reverse transcription. However; previous in vitro experiments were unable to clarify the role of the zinc fingers in this process, due to nucleic acid aggregation induced by the basic N- and C-terminal domains of NCp7. We show here that deletion of thes e sequences in (12-53)NCp7 strongly reduces the formation of aggregate s and allows a direct visualization of the binary or ternary complexes between NCp7 and nucleic acids by gel electrophoresis. (12-53)NCp7 is able to induce hybridization of the P-33 tRNA(3)(Lys) and the human i mmunodeficiency virus type 1 viral RNA-(77-257), which contains the pr imer binding site. Modification of the proximal zinc finger conformati on in Cys(23)(12-53)NCp7 led to a large reduction in this hybridizatio n process, while replacement of Trp(37) by Leu in the distal zinc fing ers resulted in a complete absence of annealing activity. These data a ccount for the in vivo loss of viral infectivity following these mutat ions and emphasize the critical role of the structure of the zinc fing er domain of NCp7. This could facilitate a rational approach to new an tiviral agents directed toward NCp7.