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Identification of tRNAs incorporated into wild-type and mutant human immunodeficiency virus type 1

Jiang, Min, Mak, Johnson, Ladha, Azim, Cohen, Eric, Klein, Michel, Rovinski, Benjamin and Kleiman, Lawrence 1993, Identification of tRNAs incorporated into wild-type and mutant human immunodeficiency virus type 1, Journal of virology, vol. 67, no. 6, pp. 3246-3253.

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Title Identification of tRNAs incorporated into wild-type and mutant human immunodeficiency virus type 1
Author(s) Jiang, Min
Mak, Johnson
Ladha, Azim
Cohen, Eric
Klein, Michel
Rovinski, Benjamin
Kleiman, Lawrence
Journal name Journal of virology
Volume number 67
Issue number 6
Start page 3246
End page 3253
Total pages 8
Publisher American Society for Microbiology
Place of publication Washington, D. C.
Publication date 1993-06
ISSN 0022-538X
1098-5514
Keyword(s) tRNA
HIV-1IIIB
DNA
RNA
wild-type virus
viral
Summary We have identified the tRNAs which are incorporated into both wild-type human immunodeficiency virus type 1 strain IIIB (HIV-1IIIB) produced in COS-7 cells transfected with HIV-1 proviral DNA and mutant, noninfectious HIV-1Lai particles produced in a genetically engineered Vero cell line. The mutant proviral DNA contains nucleotides 678 to 8944; i.e., both long terminal repeats and the primer binding site are absent. As analyzed by two-dimensional polyacrylamide gel electrophoresis, both mutant and wild-type HIV-1 contain four major-abundance tRNA species, which include tRNA(1,2Lys), tRNA(3Lys) (the putative primer for HIV-1 reverse transcriptase) and tRNA(Ile). Identification was accomplished by comparing the electrophoretic mobilities and RNase T1 digests with those of tRNA(3Lys) and tRNA(1,2Lys) purified from human placenta and comparing the partial nucleotide sequence at the 3' end of each viral tRNA species with published tRNA sequences. Thus, the absence of the primer binding site in the mutant virus does not affect tRNA(Lys) incorporation into HIV-1. However, only the wild-type virus contains tRNA(3Lys) tightly associated with the viral RNA genome. The identification of the tightly associated tRNA as tRNA(3Lys) is based upon an electrophoretic mobility identical to that of tRNA(3Lys) and the ability of this RNA to hybridize with a tRNA(3Lys)-specific DNA probe. In addition to the four wild-type tRNA species, the mutant HIV-1-like particle contains two tRNA(His) species and three tRNA-sized species that we have been unable to identify. Their absence in wild-type virus makes it unlikely that they are required for viral infectivity.
Language eng
Field of Research 119999 Medical and Health Sciences not elsewhere classified
Socio Economic Objective 970111 Expanding Knowledge in the Medical and Health Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©1993, American Society for Microbiology
Persistent URL http://hdl.handle.net/10536/DRO/DU:30047524

Document type: Journal Article
Collections: School of Medicine
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