Incorporation of tRNA into normal and mutant HIV-1

Kleiman, Lawrence; Caudry, S. D.; Boulerice, F; Wainberg, Mark A.; Parniak, Michael A.

doi:10.1016/0006-291x(91)91559-u

Cited by 25 publications

(20 citation statements)

References 26 publications

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“…tRNA 3 Lys is the natural primer of all immunodeficiency viruses, including the type 1 human immunodeficiency virus (HIV-1) [1][2][3]. The HIV-1 nucleocapsid protein (NC) is a short, basic, nucleic-acid binding protein with two zinc finger domains, each containing the invariant CCHC metal-ion binding motif.…”

Section: Introductionmentioning

confidence: 99%

New insights into the formation of HIV-1 reverse transcription initiation complex

et al. 2007

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HIV-1 reverse transcriptase uses the host tRNA 3 Lys as a primer for the synthesis of the minus DNA strand. The first event in viral replication is thus the annealing of tRNA to the primer binding site (PBS) in the 5' UTR of the viral RNA. This event requires a major RNA rearrangement which is chaperoned by the viral NC protein. The binding of NC to nucleic acids is essentially non-specific, however, NC is known to bind selectively to hairpins located in the 5' region of the viral RNA. In a previous study, using an NMR approach in which the reaction is slowed down by controlling temperature, we were able to follow details in this RNA unfolding/refolding process and to uncover an intermediate state. We showed that annealing initiates at the junction between the acceptor and the TΨC stems, and that, at physiological temperature, complete annealing is reached only in the presence of NC, probably when the zinc fingers contact the TΨC/D loops.In the present work, we have refined our model of the formation of the tRNA 3 Lys /PBS duplex. First, we show that annealing can initiate both from the single-stranded CCA 3'-end bases of the acceptor stem and from the bases in the TΨC stem. Secondly, by NMR and fluorescence spectroscopy, we have studied the complex between the NC protein and RNA hairpins that mimic the D and T-arms of the tRNA 3 Lys . Interestingly, the NC protein shows strong and specific binding to the D-arm of tRNA 3 Lys , which could explain the overall annealing mechanism.

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Section: Introductionmentioning

confidence: 99%

New insights into the formation of HIV-1 reverse transcription initiation complex

et al. 2007

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“…The human immunodeficiency virus (HIV) contains two genomic RNA strands in its core, in association with the viral reverse transcriptase (RT) enzyme (Ferris et al, 1990;Lowe et al, 1988;Restle et al, 1990) and primer tRNA lys~ (Jiang et al, 1992;Kleiman et al, 1991). Reverse transcription of the genomic RNA template to form proviral dsDNA is thought to occur in the host cell cytoplasm after core entry (Weiss et al, 1985).…”

Section: Introductionmentioning

confidence: 99%

DNA found in human immunodeficiency virus type 1 particles may not be required for infectivity

Arts¹,

Mak²,

Kleiman³

et al. 1994

Journal of General Virology

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We have studied the presence and significance of retroviral genome-derived DNA in the core of human immunodeficiency virus (HIV) particles produced from transfections of HXB2 expression vectors in COS-7 cells and from HIV type 1 IIIB chronically infected H9 cells. Viruses purified by sucrose cushion centrifugation and treated with DNase I contained 1000-fold more viral RNA than DNA. However protease-defective viruses that contained only pl60 ga~p°z had less than 100 times the amount of DNA in their cores than wild-type viruses suggesting that the p66/p51 form of reverse transcriptase was responsible for DNA transcription. Viruses produced by transfections in the presence of 3'-azido-3'-deoxythymidine (AZT) contained the viral RNA genome but only DNA of premature length because of the chain terminating effects of AZT. However such viruses were as infectious for CD4 + cells as wild-type virus. We conclude that retrovirus-derived DNA in HIV-1 partides is not required for infection and does not play a significant role in this process.

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“…Human immunodeficiency virus, type 1 (HIV-1) uses human tRNA 3 Lys , one of three major tRN Lys isoacceptors, for the priming of reverse transcription (1). tRNA 3 Lys is selectively incorporated into the virus particle and is annealed via 18 bases at its 3Ј-end to a complementary 18-base sequence near the 5Ј-end of viral genomic RNA termed the primer binding site (PBS) (2)(3)(4). Once annealed, tRNA 3 Lys primes the synthesis of minus strand cDNA.…”

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confidence: 99%

A HIV-1 Minimal Gag Protein Is Superior to Nucleocapsid at in Vitro tRNA3Lys Annealing and Exhibits Multimerization-induced Inhibition of Reverse Transcription

Roldán

Warren

Russell

et al. 2005

Journal of Biological Chemistry

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HIV-1 uses tRNA 3Lys to prime reverse transcription of its viral RNA. In this process, the 3-end of tRNA 3 Lys must be annealed to the primer binding site of HIV-1 genomic RNA, and the two molecules together form a complex structure. During annealing, the nucleocapsid (NC) protein enhances the unwinding of tertiary structures within both RNA molecules. Moreover, the packaging of tRNA 3Lys occurs prior to viral budding at a time when NC is still part of the Pr55Gag polyprotein. In contrast, Pr55Gag is able to produce virus-like particles on its own. We have recently shown that an N-terminal extended form of NC (mGag), containing all of the minimal elements required for virus-like particle formation, possesses greater affinity for HIV-1 genomic RNA than does NC alone. We have now studied the tRNA 3 Lys -annealing properties of mGag in comparison to those of NC and report that the former is more efficient in this regard than the latter. We have also tested each of a mutant version of mGag, an extended form of mGag, and an almost full-length form of Gag, and showed that all of these possessed greater tRNA-annealing capacity than did the viral NC protein. Yet, surprisingly, multimerization of Gag-related proteins did not abrogate this annealing process but rather resulted in dramatically reduced levels of reverse transcriptase processivity. These results suggest that the initial stages of reverse transcription may be regulated by the multimerization of Pr55Gag polyprotein at times prior to the cleavage of NC.

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Incorporation of tRNA into normal and mutant HIV-1

Cited by 25 publications

References 26 publications

New insights into the formation of HIV-1 reverse transcription initiation complex

New insights into the formation of HIV-1 reverse transcription initiation complex

DNA found in human immunodeficiency virus type 1 particles may not be required for infectivity

A HIV-1 Minimal Gag Protein Is Superior to Nucleocapsid at in Vitro tRNA3Lys Annealing and Exhibits Multimerization-induced Inhibition of Reverse Transcription

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