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Query: UMLS:C0019693 (HIV)
 170,526 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

3'-Azido-3'-deoxythymidine triphosphate (erythro) and its threo isomer were synthesized and investigated for their inhibition of HIV reverse transcriptase from virus isolate U 937/HTLV-III. The erythro isomer was a competitive inhibitor of the reaction directed by (rA)n(dT)12-18 and the Ki value was 0.0022 microM. The threo isomer was at least 100-fold less active. The inhibition was specific for dTMP incorporation, and dGMP incorporation using (rC)n(dG)12-18 as template/primer was not affected. The Km value for dTTP varied between 0.7 microM and 1.7 microM. Reverse transcriptase from nine HIV isolates were tested for inhibition by the erythro isomer and only slight differences in sensitivity were observed.
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PMID:Inhibition of the reverse transcriptase from HIV by 3'-azido-3'-deoxythymidine triphosphate and its threo analogue. 244 Mar 80

Based on precedents from other retroviruses, the precursor of the human immunodeficiency virus (HIV-1) reverse transcriptase is predicted to be a polyprotein with a relative molecular mass (Mr) of 160,000 (160K) encoded by both the viral pol gene and the upstream gag gene. These two genes lie in different translational reading frames, with the 3' end of gag overlapping the 5' end of pol by 205 or 241 nucleotides. Thus, production of the gag-pol fusion protein would require either messenger RNA processing or translational frameshifting. The latter mechanism has been shown in the synthesis of the gag-pol proteins of two other retroviruses, Rous sarcoma virus (RSV) and mouse mammary tumour virus (MMTV). Here we report that translation of HIV-1 RNA synthesized in vitro by SP6 RNA polymerase yields significant amounts of a gag-pol fusion protein, indicating that efficient ribosomal frameshifting also occurs within the HIV-1 gag-pol overlap region. Site-directed mutagenesis and amino-acid sequencing localized the site of frameshifting to a UUA leucine codon near the 5' end of the overlap.
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PMID:Characterization of ribosomal frameshifting in HIV-1 gag-pol expression. 244 6

In vitro experiments were conducted to assess whether bedbugs (Cimex lectularius and Cimex hemipterus) and mosquitoes (Aedes aegypti formosus) could act as vectors of HIV. These insects engorged through a membrane on a blood-virus mixture. Female bedbugs were larger than males and took larger blood-meals when fed to repletion. It was determined that the full blood-meal of a female bedbug contained 0.09 x 10(5) tissue culture infectious doses (TCID) of virus and a male 0.07 x 10(5) TCID, while partial meals taken when feeding was interrupted contained 0.013 x 10(5) TCID and 0.015 x 10(5) TCID for female and male bugs, respectively. Reverse transcriptase activity was assayed after culture of insect extracts in H9 cells: this showed survival of virus in C. lectularius for up to 4 h, in C. hemipterus for up to 1, possibly 2 h, but no survival in Ae. aegypti formosus. Four attempts to transmit the virus by interrupted feeding by C. lectularius from a blood-virus mixture to uninfected blood failed. It is concluded that Ae. aegypti formosus and probably other mosquitoes are not mechanical vectors of HIV and that such transmission is also unlikely to occur in bedbugs under natural conditions.
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PMID:Experimental assessment of bedbugs (Cimex lectularius and Cimex hemipterus) and mosquitoes (Aedes aegypti formosus) as vectors of human immunodeficiency virus. 245 May 52

The human immunodeficiency virus type 1 (HIV-1) shows extensive genetic variation and undergoes rapid evolution. The fidelity of purified HIV-1 reverse transcriptase was measured during DNA polymerization in vitro by means of three different assays. Reverse transcriptase from HIV-1 introduced base-substitution errors in DNA from the bacteriophage phi X174 amber3 at estimated frequencies of 1/2000 to 1/4000. Analyses of misincorporation rates opposite a single template adenine residue showed that HIV-1 reverse transcriptase catalyzed nucleotide mismatches with a specificity of A:C much greater than A:G greater than A:A. The high error rate of HIV-1 reverse transcriptase in vitro translates to approximately five to ten errors per HIV-1 genome per round of replication in vivo. This high error rate suggests that misincorporation by HIV-1 reverse transcriptase is, at least in part, responsible for the hypermutability of the AIDS virus. The specificity of misincorporation may provide a basis for the systematic construction of antiviral nucleosides.
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PMID:Fidelity of HIV-1 reverse transcriptase. 246 Sep 24

Reverse transcriptase from the human immunodeficiency virus type I (HIV-1) was expressed in E. coli and purified to near homogeneity. The enzyme was shown to contain reverse transcriptase, DNA polymerase and ribonuclease H activities. The DNA polymerase activity converted singly-primed phi X174 (+) DNA into the double-stranded form. Two third of the replication product is ligatable to covalently closed circular DNA (RFIV-form DNA) indicating that DNA synthesis by HIV reverse transcriptase can proceed until the enzyme matches the 5'-end of a pre-existing primer molecule. The in vitro accuracy of HIV reverse transcriptase was measured with the phi X174am16 reversion assay to be 1/7,400. Reversion rates for the individual mispairs were determined from pool bias studies to be 1/8,000 for the dGMP:T template mismatch, 1/35,000 for the dGMP:A template mismatch, 1/45,000 for the dAMP:G template mismatch, 1/73,000 for the dCMP:T template mispair, 1/140,000 for the dCMP:A template mispair, and 1/180,000 for the dGMP:G template mismatch. The dTMP:T template mispair was below the detection limit of the assay indicating a reversion rate of less than 1/300,000 for this particular mispair.
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PMID:Fidelity of human immunodeficiency virus type I reverse transcriptase in copying natural DNA. 246 38

Carbocylic 2',3'-didehydro-2',3'-dideoxyguanosine (Carbovir; NSC 614846) is an antiretroviral agent which may be useful in the treatment of AIDS. We have synthesized the 5'-triphosphate of Carbovir and examined its ability to inhibit human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (EC 2.7.7.49) and other retroviral reverse transcriptases, as well as human DNA polymerases alpha, beta, gamma (EC 2.7.7.7) and DNA primase (EC 2.7.7.6). Carbovir triphosphate emerges as a highly selective inhibitor of reverse transcriptases with little, if any, effect on the cellular enzymes. 3'-Azido-2',3'-dideoxythymidine (AZT) triphosphate and the two dideoxynucleoside triphosphates, ddTTP and ddGTP, inhibited HIV-1 reverse transcriptase to the same degree as Carbovir triphosphate, but were less selective in that they also inhibited DNA polymerases beta and gamma. We conclude that Carbovir is a highly selective antiretroviral agent.
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PMID:Comparison of the effect of Carbovir, AZT, and dideoxynucleoside triphosphates on the activity of human immunodeficiency virus reverse transcriptase and selected human polymerases. 247 36

We have dissected the protein and nucleic acid determinants that direct a group of transcriptional antiterminators to their specific target operons. These antiterminators, the N gene products of phages lambda, 21, and P22, function solely with their respective recognition sites, nut, to modify RNA polymerase to a termination-resistant form. We demonstrate that a unique hairpin sequence within each nut site, called boxB, confers genome specificity by interacting with a small amino-terminal domain of the cognate N protein. This interaction is dependent upon an arginine-rich subdomain, which is conserved not only among the N proteins but also in many RNA binding proteins from ribosomes and RNA virus capsids. Notably, this motif constitutes an essential domain of the HIV protein Tat whose function as a trans-activator requires a specific hairpin sequence.
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PMID:Sequence-specific recognition of RNA hairpins by bacteriophage antiterminators requires a conserved arginine-rich motif. 247 56

Effects of erythro-9(2-hydroxy-3-nonyl) adenine (EHNA), an inhibitor of the common Adenosine deaminase (adenosine aminohydrolase, EC 3.5.4.4.), on HIV-1 production was evaluated in vitro. Reverse transcriptase (RT) activity in the supernatant was inhibited by nearly 50% in EHNA-treated HIV-1 infected H9 cells, when compared with untreated but infected H9 cells. There was also a significant decrease in cell viability, but this was reversed following the addition of deoxycytidine (dC) to these cultures. The combined treatment was also effective in suppressing HIV-1 release from HIV-1-infected U937 cells. This combined EHNA plus dC treatment had no effect on RT activity in the cell lysates, suggesting that the inhibition of HIV-1 production may be due to the disturbance of virus release from infected cells.
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PMID:Evaluation of the effects of erythro-9(2-hydroxy-3-nonyl) adenine (EHNA) on HIV-1 production in vitro. 247 29

Trans-activation of HIV-1 transcription by the viral regulatory protein Tat has been proposed recently to overcome a block to RNA polymerase II elongation in vivo imposed by 5'-untranslated leader sequences. Interestingly, however, only full-length transcripts, rather than prematurely terminated HIV RNAs, are synthesized in most cell-free transcription extracts. Here, we describe an in vitro system in which induction of a highly efficient RNA polymerase II termination or cotranscriptional RNA processing event creates short HIV RNAs with 3' ends that map to a region immediately downstream of the HIV-1 or HIV-2 promoters. Termination in vitro is sequence dependent, generating short HIV-1 RNAs of 58-61 nucleotides that resemble in vivo transcripts observed in the absence of Tat, and a distinct, longer transcript of approximately 125-130 nucleotides from the HIV-2 promoter. Deletion of promoter-proximal HIV-2 downstream sequences results in the loss of a discrete RNA but also fails to restore wild-type transcription, indicating that termination actually is specified at the promoter and occurs at a site positioned by one or more elements located immediately upstream of the 3' end of the short RNAs. Experiments with recombinant HIV-2 promoters and nucleoside analogs indicate that this event involves a concerted interaction between the promoter and orientation-dependent leader sequences and that RNA secondary structure formation may also be required. These data provide direct evidence for abbreviated HIV transcripts and an in vitro approach to understanding the roles of cellular and viral regulatory proteins that mediate this process at the HIV promoters.
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PMID:In vitro formation of short RNA polymerase II transcripts that terminate within the HIV-1 and HIV-2 promoter-proximal downstream regions. 254 24

Promoter-specific transcription factors, whose function was once thought to be limited to initiation, are now known to have more diverse roles in RNA metabolism, including the cellular localization of transcripts and the integration of RNA initiation with attenuation and RNA 3' end formation. The human immunodeficiency viruses (HIV-1 and HIV-2) provide a useful system to study such proteins, since distinct DNA and RNA elements downstream of the site of transcription initiation act in conjunction with the promoter to regulate the induction and attenuation of RNA synthesis. Sequences corresponding to the 5' untranslated leader of HIV-1 and HIV-2 harbor at least three distinct elements: (i) a DNA domain that binds LBP-1, a cellular activator of initiation; (ii) a structured RNA element critical for the function of the HIV-1 trans-activating protein, Tat; and (iii) an RNA element required for the production of attenuated RNAs from the basal (uninduced) promoter. These attenuated leader RNAs seem to be created in vitro by stalled RNA polymerase II complexes that may be uniquely capable of rapidly processing RNA. Tat-mediated increases in steady-state levels of viral transcripts appear from nuclear run-on experiments to involve a control mechanism at both initiation and early post-initiation steps. Studies that implicate a role for Tat in post-transcriptional control suggest the existence of a mechanism for the coordination of eukaryotic transcription and translation, possibly through the assembly of nuclear regulatory factors at the 5' end of the RNA.
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PMID:HIV trans-activation and transcription control mechanisms. 256 18


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