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MOLECULAR CLONING AND ANALYSIS OF FUNCTIONAL ENVELOPE GENES FROM HUMAN IMMUNODEFICIENCY VIRUS TYPE 1 SEQUENCE SUBTYPES A THROUGH G.
HIV-1/genética
Sequência de Aminoácidos
Animais
Sequência de Bases
Clonagem Molecular
DNA Viral
Expressão Gênica/efeitos de drogas
Produtos do Gene env/fisiologia
Proteína gp120 do Envelope de HIV/genética
Proteína gp41 do Envelope de HIV/fisiologia
Infecções por HIV/virologia
HIV-1/classificação
Células HeLa
Humanos
Dados de Sequência Molecular
Filogenia
Reação em Cadeia da Polimerase
Precursores de Proteínas/genética
Recombinação Genética
Tunicamicina/farmacologia
Autor(es)
Gao, Feng
Morrison, Sandre G
Robertson, David L
Thornton, Charlotte L
Craig, Stevenson
Karlsson, Gunilla
Sodroski, Joseph
Morgado, Mariza Gonçalves
Castro Filho, Bernardo Galvão
Von Briesen, Hagen
Beddows, Simon
Weber, Jonathan
Sharp, Paul M
Shaw, George M
Hahn, Beatrice H
Who NIAID Networks for HIV isolation and characterization
Morrison, Sandre G
Robertson, David L
Thornton, Charlotte L
Craig, Stevenson
Karlsson, Gunilla
Sodroski, Joseph
Morgado, Mariza Gonçalves
Castro Filho, Bernardo Galvão
Von Briesen, Hagen
Beddows, Simon
Weber, Jonathan
Sharp, Paul M
Shaw, George M
Hahn, Beatrice H
Who NIAID Networks for HIV isolation and characterization
Afiliação
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Nottingham. Queens Medical Centre. Department of Genetics. Nottingham
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
Dana-Farber Cancer Institute. Department of Pathology. Boston, Massachusetts
Dana-Farber Cancer Institute. Department of Pathology. Boston, Massachusetts
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório Avançado de Saúde Pública. Salvador, BA, Brasil
Chemotherapeutisches Forschungsinstitut. Georg Speyer Haus. Frankfurt am Main. Germany
St. Mary’s Hospital Medical School. Department of Communicable Diseases. Imperial College. London
St. Mary’s Hospital Medical School. Department of Communicable Diseases. Imperial College. London
University of Nottingham. Queens Medical Centre. Department of Genetics. Nottingham
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Nottingham. Queens Medical Centre. Department of Genetics. Nottingham
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
Dana-Farber Cancer Institute. Department of Pathology. Boston, Massachusetts
Dana-Farber Cancer Institute. Department of Pathology. Boston, Massachusetts
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Imunologia. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Laboratório Avançado de Saúde Pública. Salvador, BA, Brasil
Chemotherapeutisches Forschungsinstitut. Georg Speyer Haus. Frankfurt am Main. Germany
St. Mary’s Hospital Medical School. Department of Communicable Diseases. Imperial College. London
St. Mary’s Hospital Medical School. Department of Communicable Diseases. Imperial College. London
University of Nottingham. Queens Medical Centre. Department of Genetics. Nottingham
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
University of Alabama at Birmingham. Departments of Medicine and Microbiology. Birmingham, Alabama
Resumo em Inglês
Present knowledge of human immunodeficiency virus type 1 (HIV-1) envelope immunobiology has been derived almost exclusively from analyses of subtype B viruses, yet such viruses represent only a minority of strains currently spreading worldwide. To generate a more representative panel of genetically diverse envelope genes, we PCR amplified, cloned, and sequenced complete gp160 coding regions of 35 primary (peripheral blood mononuclear cell-propagated) HIV-1 isolates collected at major epicenters of the current AIDS pandemic. Analysis of their deduced amino acid sequences revealed several important differences from prototypic subtype B strains, including changes in the number and distribution of cysteine residues, substantial length differences in hypervariable regions, and premature truncations in the gp41 domain. Moreover, transiently expressed glycoprotein precursor molecules varied considerably in both size and carbohydrate content. Phylogenetic analyses of full-length env sequences indicated that the panel included members of all major sequence subtypes of HIV-1 group M (clades A to G), as well as an intersubtype recombinant (F/B) from an infected individual in Brazil. In addition, all subtype E and three subtype G viruses initially classified on the basis of partial env sequences were found to cluster in subtype A in the 3' half of their gp41 coding region, suggesting that they are also recombinant. The biological activity of PCR-derived env genes was examined in a single-round virus infectivity assay. This analysis identified 20 clones, including 1 from each subtype (or recombinant), which expressed fully functional envelope glycoproteins. One of these, derived from a patient with rapid CD4 cell decline, contained an amino acid substitution in a highly conserved endocytosis signal (Y721C), as mediated virus entry with very poor efficiency, although they did not contain sequence changes predicted to alter protein function. These results indicate that the env genes of primary HIV-1 isolates collected worldwide can vary considerably in their genetic, phylogenetic, and biological properties. The panel of env constructs described here should prove valuable for future structure-function studies of naturally occurring envelope glycoproteins as well as AIDS vaccine development efforts targeted against a broader spectrum of viruses.
DeCS
Produtos do Gene env/genéticaHIV-1/genética
Sequência de Aminoácidos
Animais
Sequência de Bases
Clonagem Molecular
DNA Viral
Expressão Gênica/efeitos de drogas
Produtos do Gene env/fisiologia
Proteína gp120 do Envelope de HIV/genética
Proteína gp41 do Envelope de HIV/fisiologia
Infecções por HIV/virologia
HIV-1/classificação
Células HeLa
Humanos
Dados de Sequência Molecular
Filogenia
Reação em Cadeia da Polimerase
Precursores de Proteínas/genética
Recombinação Genética
Tunicamicina/farmacologia
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