Details

Autor(en) / Beteiligte

• Julias, John G.
• McWilliams, Mary Jane
• Sarafianos, Stefan G.
• Arnold, Edward
• Hughes, Stephen H.

Titel

Mutations in the RNase H Domain of HIV-1 Reverse Transcriptase Affect the Initiation of DNA Synthesis and the Specificity of RNase H Cleavage in vivo

Ist Teil von

• Proceedings of the National Academy of Sciences - PNAS, 2002-07, Vol.99 (14), p.9515-9520

Ort / Verlag

United States: National Academy of Sciences

Erscheinungsjahr

2002

Quelle

MEDLINE

Beschreibungen/Notizen

• Retroviral reverse transcriptases contain a DNA polymerase activity that can copy an RNA or DNA template and an RNase H activity that degrades the viral RNA genome during reverse transcription. RNase H makes both specific and nonspecific cleavages; specific cleavages are used to generate and remove the polypurine tract primer used for plus-strand DNA synthesis and to remove the tRNA primer used for minus-strand DNA synthesis. We generated mutations in an HIV-1-based vector to change amino acids in the RNase H domain that contact either the RNA and DNA strands. Some of these mutations affected the initiation of DNA synthesis, demonstrating an interdependence of the polymerase and RNase H activities of HIV-1 reverse transcription during viral DNA synthesis. The ends of the linear DNA form of the HIV-1 genome are defined by the specific RNase H cleavages that remove the plus- and minus-strand primers; these ends can be joined to form two-long-terminal repeat circles. Analysis of two-long-terminal repeat circle junctions showed that mutations in the RNase H domain affect the specificity of RNase H cleavage.