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Multiple forms of tRNA(Lys3) in HIV-1
1Lady Davis Institute for Medical Research, Jewish General Hospital, Montreal, Quebec, Canada.
Biochemical and Biophysical Research Communications
|October 14, 1996
Summary
Multiple forms of lysine-tRNA3 (tRNALys3) are found in HIV-1, differing from the single form in cells. The major viral tRNALys3 isoacceptor is also the primary primer for HIV-1 reverse transcriptase.
Area of Science:
- Molecular Biology
- Virology
- Biochemistry
Background:
- tRNALys3 serves as the primer for HIV-1 reverse transcriptase (RT).
- Cellular tRNALys3 exists in a single detectable form, while HIV-1 incorporates multiple forms.
- These viral tRNALys3 isoacceptors are present in HIV-1 produced from various infected cell lines.
Purpose of the Study:
- To investigate the heterogeneity of tRNALys3 in HIV-1.
- To identify the specific tRNALys3 isoacceptor utilized as the primer for HIV-1 reverse transcription.
Main Methods:
- RPC-5 column chromatography to separate tRNALys3 isoacceptors.
- Northern blot analysis using a tRNALys3-specific DNA probe.
- 3' end-labeling of viral tRNALys3 followed by T1 digest analysis and comparison with known tRNALys3.
Main Results:
- Multiple tRNALys3 isoacceptors were detected in HIV-1 particles, unlike the single form in host cells.
- The cytoplasmic form of tRNALys3 was identified as the major form within the virus.
- RPC-5 chromatography resolved viral tRNALys3 into distinct peaks, with peak 3 being the predominant primer for reverse transcription.
Conclusions:
- HIV-1 selectively packages multiple tRNALys3 isoacceptors.
- The major tRNALys3 isoacceptor found in HIV-1 is also the primary primer used by HIV-1 reverse transcriptase.