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HIV-1 reverse transcriptase shows no specificity for the binding of primer tRNA(Lys3)
1Lady Davis Institute for Medical Research, Sir Mortimer B. Davis, Jewish General Hospital, Montreal, Quebec, Canada.
Biochemical and Biophysical Research Communications
|August 23, 1996
Summary
Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) does not specifically bind the primer tRNA(Lys3). This suggests other factors are involved in selecting the primer tRNA for viral particle incorporation.
Area of Science:
- Biochemistry
- Virology
- Molecular Biology
Background:
- The human immunodeficiency virus type 1 (HIV-1) transcription requires a specific cellular tRNA, tRNA(Lys3), to initiate reverse transcription.
- Understanding the interaction between HIV-1 reverse transcriptase (RT) and tRNA(Lys3) is crucial for comprehending viral replication.
Purpose of the Study:
- To investigate the binding specificity of recombinant HIV-1 RT to the primer tRNA(Lys3).
- To determine if HIV-1 RT possesses a selective binding mechanism for primer tRNA.
Main Methods:
- Gel retardation analysis
- Intrinsic RT protein fluorescence quenching assays
- Nitrocellulose filter binding assays
Main Results:
- HIV-1 RT binding to tRNA was saturable, indicating specific binding sites.
- However, RT exhibited non-selective binding, with all tRNA isoacceptors and unfractionated tRNA showing similar affinity to primer tRNA(Lys3).
- No significant binding of tRNA to RT was observed.
Conclusions:
- HIV-1 RT lacks specificity in binding primer tRNA(Lys3).
- The selective incorporation of primer tRNA into HIV-1 virions likely involves factors beyond RT binding specificity.