Tat is required for efficient HIV-1 reverse transcription

• Published in: The EMBO journal Vol. 16; no. 6; pp. 1224 - 1235
• Main Authors: Harrich, David, Ulich, Catherine, García-Martínez, León F., Gaynor, Richard B.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 17.03.1997
• Subjects: AIDS/HIV; Amino Acid Sequence; Base Sequence; Cell Line; Defective Viruses - genetics; DNA Primers - genetics; DNA, Viral - genetics; Gene Deletion; Gene Expression Regulation, Viral; Gene Products, tat - genetics; Gene Products, tat - metabolism; Genes, tat; Genetic Complementation Test; HIV-1; HIV-1 - genetics; HIV-1 - metabolism; Humans; Molecular Sequence Data; reverse transcription; tat Gene Products, Human Immunodeficiency Virus; Tat protein; Transcription, Genetic; Transfection; Virus Replication - genetics
• ISSN: 0261-4189; 1460-2075; 1460-2075
• DOI: 10.1093/emboj/16.6.1224

The ability of human immunodeficiency virus‐1 (HIV‐1) to undergo efficient reverse transcription is dependent on a number of parameters. These include the binding of the tRNA3Lys to the HIV‐1 primer binding site and the subsequent interaction with the heterodimeric reverse transcriptase. Recently, we demonstrated that TAR RNA was also necessary for efficient HIV‐1 reverse transcription. Given the fact that the Tat protein is involved in the activation of HIV‐1 gene expression in conjunction with TAR, we wished to determine whether Tat might also be involved in the control of HIV‐1 reverse transcription. HIV‐1 virions deleted in the tat gene were unable to initiate reverse transcription efficiently upon infection of peripheral blood mononuclear cells (PBMCs). This defect was not due to decreased amounts of genomic RNA, reverse transcriptase or other HIV‐1 proteins which were incorporated into the virion. Following transfection of wild‐type but not mutant tat genes into cell lines producing HIV‐1 lacking tat, the virions produced could be complemented for defects in reverse transcription upon subsequent infection of PBMCs. In contrast, the defect in reverse transcription seen with HIV‐1 lacking the tat gene could not be complemented when the target cells rather than the producer cells contained tat. Viruses lacking tat were also defective in endogenous assays of reverse transcription, although these viruses contained similar levels of reverse transcriptase. These results indicate that the Tat protein, in addition to regulating the level of gene expression, is also important for efficient HIV‐1 reverse transcription.