Synthetic human tRNAuuuLys3 and natural bovine tRNAsuuLys3 interact with HIV-1 reverse transcriptase and serve as specific primers for retroviral cDNA synthesis

Weiß, Sabine; König, Bernhard; Müller, Hans‐Joachim; Seidel, H.; Goody, Roger S.

doi:10.1016/0378-1119(92)90686-j

Cited by 30 publications

(22 citation statements)

References 26 publications

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“…This region defines the 3' boundary of the long terminal repeat (LTR) and is the called primer binding site (PBS). The events leading to initiation of the minus-strand DMA synthesis include (1) specific incorporation of the host tRNA primer into the viral particle, (2) partial unwinding of the tRNA structure to expose 18 or more nucleotides of the 3 1 end of the tRNA, (3) unwinding of the viral RNA template structure, and (4) annealing of the tRNA to the PBS or to regions located close to the PBS (Aiyar et al, 1992;Weiss et a/.,1992).…”

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

“…HIV-1 RT has been shown to preferentially bind to tRNA Lys>3 even in the presence of a 100-fold excess of other tRNAs (Barat et al, 1989). Weiss et al demonstrated that human tRNA Lys · 3 and bovine tRNA Lys ' 3 can interact with HIV-1 RT (Weiss et al, 1992). In vitro, HIV-1 RT and the tRNA Lys · 3 primer are sufficient for initiation of DNA synthesis at the PBS Weiss et al, 1992).…”

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

“…Weiss et al demonstrated that human tRNA Lys · 3 and bovine tRNA Lys ' 3 can interact with HIV-1 RT (Weiss et al, 1992). In vitro, HIV-1 RT and the tRNA Lys · 3 primer are sufficient for initiation of DNA synthesis at the PBS Weiss et al, 1992). cDNA product analysis of the viral genome indicated the presence of at least 18 nucleotides of the 3' ends of human and bovine tRNA Lys>3 .…”

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

“…cDNA product analysis of the viral genome indicated the presence of at least 18 nucleotides of the 3' ends of human and bovine tRNA Lys>3 . The 5 1 ends of both human and bovine primer molecules are removed by the RNase H activity of HIV-1 RT (Weiss et al, 1992). This suggests that RT is able to bind and unwind its tRNA primer.…”

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

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Review

1996

Biological Chemistry Hoppe-Seyler

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Proteolytic enzymes have many physiological functions in plants, bacteria, viruses, protozoa and mammals. They play a role in processes such as food digestion, complement activation or blood coagulation. The action of proteolytic enzymes is biologically controlled by proteinase inhibitors and increasing attention is being paid to the physiological significance of these natural inhibitors in pathological processes. The reason for this growing interest is that uncontrolled proteolysis can lead to irreversible damage e.g. in chronic inflammation or tumor metastasis. This review focusses on the possible role of the cystatins, natural and specific inhibitors of the cysteine proteinases, in pathological processes. Key words: Cystatins / Cysteine proteinases / Inflammation / Inhibitors / Pathological processes. Cysteine ProteinasesCysteine Proteinases, synonymous with thiol proteinases, are small proteins with molecular masses varying from 23 to 24 kDa and with two catalytic residues, Cys25 and His159, which are involved in the hydrolytic reaction. Cysteine proteinases catalyze the hydrolysis of various polypeptide substrates and are most active under reducing and mildly acidic conditions (pH 5 -6.5). They have been detected in and isolated from a large number of biological sources including plants, bacteria, animals and humans. Bacterial cysteine proteinases are produced by Clostridium histolyticum, named clostripain, by hemolytic group A streptococci and by the pathogenic anaerobic Porphyromonas gingivalis, a bacterium associated with periodontitis. Bacterial cysteine proteinases probably play a role in the penetration of normal tissues by the bacteria and in the food digestion. Viral cysteine proteinases from picornaviruses, e.g. the poliovirus and rhinovirus type I, are involved in proteolytic cleavage of precursor proteins for virus replication and for the production of new virus particles. A virus-coded cysteine proteinase is involved in this process (Kay and Dunn, 1990; Körant et a/., 1985;. HIV-I also needs proteolytic processing by cysteine proteinases to regulate the expression of viral proteins (Guy ef a/., 1991). Protozoal cysteine proteinases are produced by a number of protozoan parasites to facilitate host invasion, metabolize host proteins, to degrade the host immune molecules or to use them for intracellular replication. Cysteine proteinases are produced by e.g. Trypanosoma cruzi, the causative agent of American trypanosomiasis or by Leishmania species, causing one of the six major parasitic diseases. Furthermore, human malarial parasites produce a broad scala of proteinases including cysteine proteinase which are involved in erythrocyte invasion and rupture (McKerrow, 1993). Mammalian cysteine proteinases are present in the lysosomes of cells. Lysosomes contain different cysteine proteinases, the most important being the cathepsins B, H, L and S Kirschke, 1981, Barrett et a/., 1988). These cathepsins have common ancestors and are related to papain. Cathepsin B has been detected in every tissue or cel...

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Section: Minus-strand Dna Synthesismentioning

confidence: 99%

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

Section: Minus-strand Dna Synthesismentioning

confidence: 99%

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Review

1996

Biological Chemistry Hoppe-Seyler

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“…A powerful substance with chiral structure and high specificity for HIV-1 was identified; this substance was BM 21.1298, a thiazolo-iso-indolinone ( …”

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confidence: 99%

Viral resistance to the thiazolo-iso-indolinones, a new class of nonnucleoside inhibitors of human immunodeficiency virus type 1 reverse transcriptase

Maaß¹,

Immendoerfer²,

Koenig³

et al. 1993

Antimicrob Agents Chemother

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Thjazolo-iso-indolinone derivatives with high specificity toward the reverse transcriptase (RT) of human immunodeficiency virus type 1 (HIV-1) were identified. The most potent compound, BM +51.0836, inhibited HIV-1 RT at a 50%o inhibitory concentration of 90 nM in vitro. In cell culture assays, similar 50%o inhibitory concentrations were obtained with high specificity for HIV-1. These substances were equally active against a zidovudine-resistant isolate. No antiviral effect was observed with an HIV-2 isolate. HIV-1 isolates resistant to the thiazolo-iso-indolinones were generated in cell culture, and the nucleotide sequences of the respective RT genes were analyzed subsequently. Comparison Fig. lb], a substance with 10-fold lower 50% inhibitory concentrations (IC50s) than the lead compound BM 21.1298 and higher antiviral activity than 0-TIBO and nevirapine (BI-RG-587). MATERIALS AND METHODSCompounds. The thiazolo-iso-indolinones were synthesized by Boehringer Mannheim GmbH. A detailed description of the synthesis will be published elsewhere. Zidovudine (AZT; Wellcome), nevirapine (BI-RG-587; Boehringer Ingelheim), and 0-TIBO (Janssen) were also synthesized by the chemical department of Boehringer Mannheim GmbH. The inhibitors were dissolved in dimethyl sulfoxide, and the mixture was added to the RT assay mixture (the dimethyl sulfoxide concentration was less than 5%). For cell culture assays, stock solutions were prepared by dissolving the inhibitors in 50% dimethyl sulfoxide-50% culture medium. In the cell culture supernatant, the dimethyl sulfoxide concentration was less than 1%.Nonradioactive Bl-1 RT assay. Selective inhibitors of HIV-1 RT were identified by using a previously published high-efficiency screening system (4). The assay contains purified HIV-1 RT (11) expressed in Escherichia coli, an in vitro transcript of the HIV-1 long terminal repeat (21), and the primer-binding site as the template and as well as an 18-mer oligonucleotide as the primer. Inhibition of RT was determined by a nonradioactive RT assay (ELISA) with biotin-and digoxigenin-labeled nucleoside triphosphates (4). In brief, 1 ,ug

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