Human retroviruses and AIDS 1994

Myers, Gerald; Korber, Bette T.; Wain‐Hobson, Simon; Jeang, Kuan‐Teh; Henderson, Louis E.; Pavlakis, George N.

doi:10.2172/10116202

Cited by 246 publications

(329 citation statements)

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“…This finding is in contrast to our data, which indicate that a peptide of 55 amino acids containing both zinc-binding motifs is the minimal amino acid sequence required for the dimerization. Furthermore, neither an N-terminal peptide (positions [1][2][3][4][5][6][7][8][9][10][11][12][13][14] nor a C-terminal 39 amino acid-containing peptide (positions 13-51) (nor mixtures of both) appears to be functional. The NCplO of MoMuLV and that of RaMuLV also have been synthesized (22,23).…”

Section: Resultsmentioning

confidence: 99%

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Identification of a binding site for the human immunodeficiency virus type 1 nucleocapsid protein

Sakaguchi¹,

Zambrano²,

Baldwin³

et al. 1994

Peptides

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The nucleocapsid (NC) protein NCp7 of human immunodeficiency virus type 1 (HIV-1) is important for encapsidation of the virus genome, RNA dimerization, and primer tRNA annealing in vitro. Here we present evidence from gel mobility-shift experiments indicating that NCp7 binds specifically to an RNA sequence. Two complexes were identified in native gels. The more slowly migrating complex contained two RNA molecules and one peptide, while the more rapidly migrating one is composed ofone RNA and one peptide. Further, mutational analysis of the RNA shows that the predicted stem and loop structure ofstem-oop 1 plays a critical role. Our results show that NCp7 binds to a unique RNA structure within the i region; in addition, this structure is necessary for RNA dimerization. We propose that NCp7 binds to the RNA via a direct interaction of one zinc-binding motif to stem-oop 1 followed by binding of the other zinc-binding motif to stem-loop 1, stem-oop 2, or the linker region of the second RNA molecule, forming a bridge between the two RNAs.Type C retroviruses typically contain two copies of the single-stranded RNA genome in each virion (1). These two unspliced RNA molecules are linked together near their 5' ends by the dimer linkage structure (DLS). A specific cellular tRNA is annealed to each RNA at the primer binding site and serves as a primer during the reverse transcription of the RNA genome into DNA. Mutational analyses have shown that the assembly ofthe DLS is important for packaging ofthe viral-specific RNA. Packaging defects can be produced with genetic deletions within the q/ region, located between the lysine tRNA primer binding site and the start of the gag polyprotein coding sequences, or by mutations of codons within the gag gene (2, 3).The human immunodeficiency virus type 1 (HIV-1) nucleo- The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.In the case of HIV-1, the interaction of NC with RNA was examined by using the crosslinking reagent trans-diamminedichloroplatinum(II). A 10-nucleotide (nt) fragment was identified as the NC binding site (9). Mutations in the zinc fingers or the flanking basic residues were shown to abrogate the high-affinity RNA binding in vitro as well as the RNA annealing activities of the NC proteins (6).In this study, we report that a 44-nt RNA segment containing the HIV-1 q, region is folded into two stem-loop structures separated by a stretch of 11 nt. The 44-nt RNA oligomer (44-mer) contains the NCp7 binding site, which requires an intact stem and loop structure. The functional importance of this binding site for RNA dimerization is illustrated by a model for the DLS. MATERIALS AND METHODSPrediction of RNA Folding of HIV-1 Sequences. The sequences of eight variants of HIV-1 (HIVRF, HIVHAN, HIVJRCSF, HIVSF2, HIVLA1, HIVHXB2R, HIVNL43, HIVNY5) were aligned according to the Los Alamos convention (10), and their second...

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Section: Resultsmentioning

confidence: 99%

“…The sequences of eight variants of HIV-1 (HIVRF, HIVHAN, HIVJRCSF, HIVSF2, HIVLA1, HIVHXB2R, HIVNL43, HIVNY5) were aligned according to the Los Alamos convention (10), and their secondary structures were predicted. Each sequence consisted of300 bases symmetrically disposed about the q,site and were folded independently.…”

Section: Methodsmentioning

confidence: 99%

Identification of a binding site for the human immunodeficiency virus type 1 nucleocapsid protein

Sakaguchi¹,

Zambrano²,

Baldwin³

et al. 1994

Peptides

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“…The V3 loop of different HIV-1 and HIV-2 isolates contains a highly conserved RP dipeptide motif at its NH 2 -terminal end (6). In view of this, we have designed and synthesized a construction referred to as template assembled synthetic peptide (TASP) in which templates such as KKKGPKEKGC and KKKKGC were employed to covalently anchor arrays of tripeptides (RPR, RPK, or KPR) at the ⑀-amino group of the lysine residues in the templates (7).…”

Section: Hiv 1 Is An Enveloped Virus That Infects Target Cells By Thementioning

confidence: 99%

Pseudopeptide TASP Inhibitors of HIV Entry Bind Specifically to a 95-kDa Cell Surface Protein

Callebaut

Jacotot

Krust

et al. 1997

Journal of Biological Chemistry

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“…The human immunodeficiency virus (HIV) and other retroviruses exhibit extensive genome variability, with single-base differences, short deletions and insertions making up the majority of sequence variation (1,2). The reported rate at which mutations are generated ranges from 10-6 to 10-4 mutations per nucleotide per cycle through a host cell (3,4).…”

Section: Introductionmentioning

confidence: 99%

Efficient extension of a misaligned tRNA-primer during replication of the HIV-1 retrovirus

Das

Berkhout

1995

Nucl Acids Res

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The human immunodeficiency virus (HIV) and other retroviruses show extensive genomic variation, which is primarily due to error-prone replication by the viral reverse transcriptase (RT) enzymes. RT errors include misincorporation with subsequent extension of the mismatched terminal base, and extension of realigned primer-template duplexes. Whereas both RT-mediated mechanisms have been extensively studied in vitro, almost no in vivo experiments have been performed. In this work, we analyzed the ability of HIV-1 RT to extend a misaligned tRNALYs3 primer in vivo. This tRNA binds with its 3'-terminal 18 nt to a complementary sequence in the viral genome, referred to as the primer-binding site (PBS). We constructed a series of mutant viral genomes with small insertions or deletions in the PBS sequence, resulting in misalignment of the tRNA primer. Extension of the misaligned primer did occur with reasonable efficiency for some of the mutants, resulting in reversion to the wild-type viral sequence. The infectivity and reversion frequency of the PBS mutants is therefore a measure of the efficiency of extending a misaligned primer in vivo. Using virionderived primer-template complexes, we also measured the tRNA-priming efficiency in vitro. The combined results show that HIV-1 RT can elongate a misaligned primer and that the efficiency of primer extension is determined by the extent of the mismatch.

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Human retroviruses and AIDS 1994

Cited by 246 publications

References 0 publications

Identification of a binding site for the human immunodeficiency virus type 1 nucleocapsid protein

Identification of a binding site for the human immunodeficiency virus type 1 nucleocapsid protein

Pseudopeptide TASP Inhibitors of HIV Entry Bind Specifically to a 95-kDa Cell Surface Protein

Efficient extension of a misaligned tRNA-primer during replication of the HIV-1 retrovirus

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