3'-orf and sor genes of human immunodeficiency virus: in vitro transcription-translation and immunoreactive domains.

Arya, Suresh K.

doi:10.1073/pnas.84.15.5429

Cited by 18 publications

(6 citation statements)

References 25 publications

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“…Franchini et al (1987) reported similar results but found a slight increase of antibodies against vif with progression to AIDS. Ranki et al (1987) describe the presence of antibodies against vif in early HIV infection; similarily, Arya (1987) found in a limited amount of sera vif antibodies less frequently in AIDS patients than in healthy HIV carriers. This divergence may partially be explained by the fact that the recombinant vif protein used in our study represented the complete amino acid sequence of vif and that the proviral vif DNA was obtained from a different HIV clone than in some of the above-mentioned reports.…”

Section: Resultsmentioning

confidence: 99%

Antibodies to recombinant HIV-1 vif, tat, and nef proteins in human sera

Wieland

Kühn

Jassoy

et al. 1990

Med Microbiol Immunol

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The prevalence of antibodies against HIV-1 regulatory proteins in sera of HIV-infected patients from different stages of disease was investigated. HIV-1 vif, tat, and nef genes were cloned in procaryotic vectors and were expressed as MS-2 fusion proteins (vif and nef) or as a non-fusion protein (tat). These recombinant proteins were employed in immunoblot experiments. The specificity of the recognition was confirmed by competition experiments and with control sera from HIV-negative patients. Analysis of 136 serum samples revealed a high percentage of antibodies against nef, irrespective of the stage of disease. Antibodies against tat were found less frequently and increased from 16% to 40% with disease progression. Vif antibodies were detected only in a low percentage in early stages of disease, but their prevalence increased to 36% and 72% with progression of disease to AIDS-related complex and AIDS. Our data suggest that the detection of antibodies against nef may represent an additional and useful marker for the diagnosis of HIV infection, whereas the detection of vif antibodies may indicate disease progression.

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Section: Resultsmentioning

confidence: 99%

Antibodies to recombinant HIV-1 vif, tat, and nef proteins in human sera

Wieland

Kühn

Jassoy

et al. 1990

Med Microbiol Immunol

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“…The infectivity factor of human immunodeficiency virus type 1 (HIV-1), Vif, is a 23-kDa protein which modulates HIV-1 infection in cultured T-cell lines in a host celldependent manner (9,16,20,22,29,36,37) and is essential for infection of peripheral blood lymphocytes (1,7,9). Conservation of the vif open reading frame among many animal lentiviruses (23,38) and Vif immunogenicity in HIV-1-infected individuals (2,14,18,32) suggest that Vif is important in natural HIV-1 infection. Vif is both synthesized and active at a late phase of the viral life cycle (10,12,17,33), but it has no detectable effects on viral transcription or translation or virus secretion (1,7,16,20,36).…”

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confidence: 99%

Efficiency of viral DNA synthesis during infection of permissive and nonpermissive cells with vif-negative human immunodeficiency virus type 1

Sova

Volsky

1993

J Virol

159

110

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The rate-limiting steps in infection by human immunodeficiency virus type 1 (HIV-1) deficient in the viral infectivity factor, Vif, are unknown. As a measurement of completion of the early stages of the HIV-1 life cycle, the levels of viral DNA were examined by polymerase chain reaction amplification during infection by vif-positive and vif-negative viruses of MT-2 and H9 cells, in which vif is required for HIV-1 replication. Viral DNA was detected within hours of infection by both viruses, but the accumulation of vif-negative virus DNA was impeded in terms of both extent and kinetics. Inefficient viral DNA synthesis correlated with restricted replication of the vif-negative virus. Increasing the input dose of vif-negative virus increased viral DNA levels within 24 h of infection but failed to overcome the block to subsequent DNA synthesis and productive infection. Infection of C8166 cells, in which vif function is dispensable, resulted in efficient DNA synthesis by vif-positive and vif-negative viruses. We conclude that one defect in the replication of vif-negative HIV-1 in nonpermissive cells occurs prior to or during viral DNA synthesis and may reflect processes required for efficient nucleocapsid internalization or activation of reverse transcription.

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“…As with other retroviruses, full-length RNA is translated into the gag core structural proteins and also, by a translational frameshift, the pol products (12), while the env mRNA is generated by a single splicing event. There are also a number of smaller, multiply spliced transcripts present in HIV-1-infected cells which code for some of the other genes of HIV-1, including the regulatory genes tat, rev, and nef (1,2,16). These transcripts consist of three or more exons, and their expression and that of the gag-pol and env mRNAs are differentially regulated.…”

mentioning

confidence: 99%

Structure and expression of tat-, rev-, and nef-specific transcripts of human immunodeficiency virus type 1 in infected lymphocytes and macrophages

Robert-Guroff

Popovič

Gartner

et al. 1990

J Virol

211

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Primary RNA transcripts from the human immunodeficiency virus type 1 (HIV-1) are processed into mature mRNA by a complex series of splicing events. Viral structural proteins and reverse transcriptase are translated from unspliced or singly spliced transcripts. Proteins which control virus replication, including tat, rev, and nef, are translated from transcripts which are the product of multiple splicing. We have analyzed the composition and relative abundance of the latter transcripts in long-term infected cell lines and in acutely infected peripheral blood cells by amplification with the polymerase chain reaction (PCR) foUlowed by Southern blot, molecular cloning, and DNA sequence analyses. In H9 cells chronically infected with the HIV-1 strain HTLV-IIIB, the predominant of the three kinds of transcripts is those coding for nef. Transcripts with coding potential for rev constituted an intermediate fraction of those analyzed, while those for tat accounted for only a small minority. A similar pattern was observed with Southern blots of PCR-amplified transcripts from peripheral blood lymphocytes acutely infected with HTLV-IIIB. The same general pattern was also observed with PCR-amplffied transcripts from peripheral blood monocyte-macrophages infected with an HIV-1 strain (BA-L) able to grow to high titers in macrophages. In these cells, however, the apparent major form of nef transcript contained only the first and third exons of the multiply spliced transcripts and appeared to be generated by either a single or a triple splicing mechanism. As with lymphocytes, tat-specific mRNAs were by far the least abundant. It thus appears that different cell types infected with different strains of HIV-1 maintain a similar balance of expression in which transcripts for nef vastly predominate over those for tat and that those for rev are intermediate in abundance.

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3'-orf and sor genes of human immunodeficiency virus: in vitro transcription-translation and immunoreactive domains.

Abstract: An in vitro transcription and translation procedure was designed to translate multiple open reading frames from cloned DNAs. For human immunodeficiency virus (HIV) cloned DNA carrying three open reading frames (sor, tat, and 3'-orf), the approach yielded three authentic polypeptides.

Cited by 18 publications

References 25 publications

Antibodies to recombinant HIV-1 vif, tat, and nef proteins in human sera

Antibodies to recombinant HIV-1 vif, tat, and nef proteins in human sera

Efficiency of viral DNA synthesis during infection of permissive and nonpermissive cells with vif-negative human immunodeficiency virus type 1

Structure and expression of tat-, rev-, and nef-specific transcripts of human immunodeficiency virus type 1 in infected lymphocytes and macrophages

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