Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone

Adachi, Akio; Gendelman, Howard Eliot; Koenig, Scott; Folks, Thomas M.; Willey, R L; Rabson, Arnold B.; Martin, Malcom A.

doi:10.1128/jvi.59.2.284-291.1986

Cited by 2,732 publications

(828 citation statements)

References 38 publications

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“…The sequence of NC and the nucleic acids used in this study were derived from the HIV-1 pNL4-3 clone (GenBank accession no. AF324493) (Adachi et al, 1986). Sequences of the purified Gag and Gag cleavage proteins were derived from HIV-1 BH10 (GenBank accession no.…”

Section: Methodsmentioning

confidence: 99%

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Fundamental differences between the nucleic acid chaperone activities of HIV-1 nucleocapsid protein and Gag or Gag-derived proteins: Biological implications

Datta

Mitra

et al. 2010

Virology

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The HIV-1 Gag polyprotein precursor has multiple domains including nucleocapsid (NC). Although mature NC and NC embedded in Gag are nucleic acid chaperones (proteins that remodel nucleic acid structure), few studies include detailed analysis of the chaperone activity of partially processed Gag proteins and comparison with NC and Gag. Here we address this issue by using a reconstituted minus-strand transfer system. NC and NC-containing Gag proteins exhibited annealing and duplex destabilizing activities required for strand transfer. Surprisingly, unlike NC, with increasing concentrations, Gag proteins drastically inhibited the DNA elongation step. This result is consistent with “nucleic acid-driven multimerization” of Gag and the reported slow dissociation of Gag from bound nucleic acid, which prevent reverse transcriptase from traversing the template (“roadblock” mechanism). Our findings illustrate one reason why NC (and not Gag) has evolved as a critical cofactor in reverse transcription, a paradigm that might also extend to other retrovirus systems.

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Section: Methodsmentioning

confidence: 99%

“…The PR mutant, PR D25G , was a generous gift from David Ott, SAIC-Frederick, Inc., NCI-Frederick, Frederick, MD (Ott et al, 2000). The WT HIV-1 plasmid pNL4-3 (Adachi et al, 1986) was obtained from Malcolm Martin (NIAID, NIH) through the AIDS Research and Reference Reagent Program, Division of AIDS, NIAID, NIH.…”

Section: Virus Productionmentioning

confidence: 99%

Fundamental differences between the nucleic acid chaperone activities of HIV-1 nucleocapsid protein and Gag or Gag-derived proteins: Biological implications

Datta

Mitra

et al. 2010

Virology

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“…HEK 293T cells sustain all viral steps with the exception of virion attachment and entry as they lack cell receptors recognized by HIV-1. They can be transfected with the HIV-1 proviral genome to produce fully infectious viral particles, indicating that their cytoplasmic/nuclear protein makeup is capable of sustaining viral transcription and replication (55).…”

Section: Reporter Assaymentioning

confidence: 99%

Structure and possible function of a G-quadruplex in the long terminal repeat of the proviral HIV-1 genome

Nicola

Lech

Heddi

et al. 2016

Nucleic Acids Research

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The long terminal repeat (LTR) of the proviral human immunodeficiency virus (HIV)-1 genome is integral to virus transcription and host cell infection. The guanine-rich U3 region within the LTR promoter, previously shown to form G-quadruplex structures, represents an attractive target to inhibit HIV transcription and replication. In this work, we report the structure of a biologically relevant G-quadruplex within the LTR promoter region of HIV-1. The guanine-rich sequence designated LTR-IV forms a well-defined structure in physiological cationic solution. The nuclear magnetic resonance (NMR) structure of this sequence reveals a parallel-stranded G-quadruplex containing a single-nucleotide thymine bulge, which participates in a conserved stacking interaction with a neighboring single-nucleotide adenine loop. Transcription analysis in a HIV-1 replication competent cell indicates that the LTR-IV region may act as a modulator of G-quadruplex formation in the LTR promoter. Consequently, the LTR-IV G-quadruplex structure presented within this work could represent a valuable target for the design of HIV therapeutics.

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“…The HIV-1 virus stock was prepared from the Jurkat cells transfected with an HIV-1 genome-encoding plasmid, pNL4-3, originally constructed from the NY5 and RAV proviruses (Adachi et al 1986). Infectivity of HIV-1 was measured with the median tissue culture infectious dose (TCID 50 ) method using Jurkat cells.…”

Section: Anti-hiv Activitymentioning

confidence: 99%

High-Mannose Specific Lectin and Its Recombinants from a Carrageenophyta Kappaphycus alvarezii Represent a Potent Anti-HIV Activity Through High-Affinity Binding to the Viral Envelope Glycoprotein gp120

et al. 2015

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We previously reported that a high-mannose binding lectin KAA-2 from the red alga Kappaphycus alvarezii, which is an economically important species and widely cultivated as a source of carrageenans, had a potent anti-influenza virus activity. In this study, the full-length sequences of two KAA isoforms, KAA-1 and KAA-2, were elucidated by a combination of peptide mapping and cDNA cloning. They consisted of four internal tandem-repeated domains, which are conserved in high-mannose specific lectins from lower organisms, including a cyanobacterium Oscillatoria agardhii and a red alga Eucheuma serra. Using an Escherichia coli expression system, an active recombinant form of KAA-1 (His-tagged rKAA-1) was successfully generated in the yield of 115 mg per a litter of culture. In a detailed oligosaccharide binding analysis by a centrifugal ultrafiltration-HPLC method with 27 pyridylaminated oligosaccharides, His-tagged rKAA-1 and rKAA-1 specifically bound to high-mannose N-glycans with an exposed α1-3 mannose in the D2 arm as the native lectin did. Predicted from oligosaccharide-binding specificity, a surface plasmon resonance analysis revealed that the recombinants exhibit strong interaction with gp120, a heavily glycosylated envelope glycoprotein of HIV with high association constants (1.48-1.61×10 9 M −1 ). Native KAAs and the recombinants inhibited the HIV-1 entry at IC 50 s of low nanomolar levels (7.3-12.9 nM). Thus, the recombinant proteins would be useful as antiviral reagents targeting the viral surface glycoproteins with high-mannose N-glycans, and the cultivated alga K. alvarezii could also be a good source of not only carrageenans but also this functional lectin(s).

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Production of acquired immunodeficiency syndrome-associated retrovirus in human and nonhuman cells transfected with an infectious molecular clone

Cited by 2,732 publications

References 38 publications

Fundamental differences between the nucleic acid chaperone activities of HIV-1 nucleocapsid protein and Gag or Gag-derived proteins: Biological implications

Fundamental differences between the nucleic acid chaperone activities of HIV-1 nucleocapsid protein and Gag or Gag-derived proteins: Biological implications

Structure and possible function of a G-quadruplex in the long terminal repeat of the proviral HIV-1 genome

High-Mannose Specific Lectin and Its Recombinants from a Carrageenophyta Kappaphycus alvarezii Represent a Potent Anti-HIV Activity Through High-Affinity Binding to the Viral Envelope Glycoprotein gp120

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