Repair of a Rev-minus human immunodeficiency virus type 1 mutant by activation of a cryptic splice site.
We isolated a revertant virus after prolonged culturing of a replication-impaired human immunodeficiency virus type 1 (HIV-1) mutant of which the Rev open reading frame was inactivated by mutation of the AUG translation initiation codon. Sequencing of the tat-rev region of this revertant virus identified a second-site mutation in tat that restored virus replication in the mutant background. This mutation activated a cryptic 5' splice site (ss) that, when used in conjunction with the regular HIV 3' ss #5, fuses the tat and rev reading frames to encode a novel T-Rev fusion protein that rescues Rev function. We also demonstrate an alternative route to indirectly activate this cryptic 5' ss by mutational inactivation of an adjacent exon splicing silencer element.
Verhoef
K
K
Department of Human Retrovirology, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands.
Bilodeau
P S
PS
van Wamel
J L
JL
Kjems
J
J
Stoltzfus
C M
CM
Berkhout
B
B
D000818:Animals; D019556:COS Cells; D003062:Codon; D015694:Gene Products, rev / Q000235:genetics*; D015756:Genes, tat; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology; D009154:Mutation; D014779:Virus Replication; D054321:rev Gene Products, Human Immunodeficiency Virus
D003062:Codon; D015694:Gene Products, rev; D054321:rev Gene Products, Human Immunodeficiency Virus