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Rigourd, M ; Goldschmidt, V ; Brule, F ; Morrow, CD ; Ehresmann, B ; Ehresmann, C ; Marquet, R

Structure-function relationships of the initiation complex of HIV-1 reverse transcription : the case of mutant viruses using tRNA(His) as primer

Nucleic Acids Research 31 (19) 5764-5775

par Administrateur - publié le

Abstract :

Reverse transcription of HIV-1 RNA is initiated from the 3’ end of a tRNA(3)(Lys) molecule annealed to the primer binding site (PBS). An additional interaction between the anticodon loop of tRNA(3)(Lys) and a viral A-rich loop is required for efficient initiation of reverse transcription of the HIV-1 MAL isolate. In the HIV-1 HXB2 isolate, simultaneous mutations of the PBS and the A-rich loop (mutant His-AC), but not of the PBS alone (mutant His) allows the virus to stably utilize tRNA(His) as primer. However, mutant His-AC selects additional mutations during cell culture, generating successively His-AC-GAC and His-AC-AT-GAC. Here, we wanted to establish direct relationships between the evolution of these mutants in cell culture, their efficiency in initiating reverse transcription and the structure of the primer/template complexes in vitro.