Inhibition of HIV-1 replication and reactivation from latency by tat transdominant negative mutants in the cysteine rich region.
Tat mutants (tat22, tat37 and tat22/37) were constructed in the transactivation domain, where cysteines at positions 22 or/and 37 were substituted with glycine and serine, respectively. These mutants were expressed either in a BK virus episomal vector or in the retroviral vector LXSN. Constitutive production of tat22 by Jurkat T cells in the context of both vectors blocked HIV-1 replication during lytic infection. Conversely, the tat37 mutant did not show any inhibitory activity and tat22/37 displayed a mild effect on HIV-1 infection only when expressed by the recombinant retrovirus. However, constitutive production of tat22/37 by the BK virus vector in Jurkat T cells chronically infected by HIV-1 was effective in blocking reactivation of viral replication induced by tumor necrosis factor-alpha or human herpesvirus-6. These results suggest that mutants in the transactivation domain of tat may be considered in designing alternative strategies to control HIV-1 replication and reactivation from latency during different phases of infection.
Caputo
A
A
Institute of Microbiology, School of Medicine, University of Ferrara, Italy.
Grossi
M P
MP
Bozzini
R
R
Rossi
C
C
Betti
M
M
Marconi
P C
PC
Barbanti-Brodano
G
G
Balboni
P G
PG
D001483:Base Sequence; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism*; D005799:Genes, Dominant; D015756:Genes, tat; D015497:HIV-1 / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D015533:Transcriptional Activation; D014775:Virus Activation / Q000235:genetics*; D017735:Virus Latency / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*
D009838:Oligodeoxyribonucleotides; D003545:Cysteine