The nef gene of simian immunodeficiency virus SIVmac1A11

Ronald E. Unger, Marta Marthas, Elissa Pratt-Lowe, Phillip A. Padrid, Paul A Luciw

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Abstract

The role of the simian immunodeficiency virus (SIV) nef gene in viral replication was investigated in several tissue culture systems. SIVmac1A11 is a molecularly cloned virus which replicates in both peripheral blood mononuclear cells (PBMC) and macrophages, although no disease is observed in infected rhesus macaques. In this report, we demonstrate that SIVmac1A11 contains a full open reading frame for nef which specifies a 37-kDa protein. To investigate the effects of nef on viral replication, a 70-bp deletion was introduced into the nef gene of SIVmac1A11. Analysis of infected cell extracts by immunoblotting revealed that both SIVmac1A11 and nef deletion virus SIVmac1A11Δnef produced the same viral proteins, except that Nef was absent in the mutant virus. The deletion mutation did not affect viral replication in PBMC, in monocyte-derived and alveolar macrophages obtained from rhesus macaques, and in human cell lines HUT-78 and CEMx-174. In addition, SIVmac1A11 and SIVmac1A11Δnef exhibited similar patterns of cytopathologic changes and ultrastructural appearances in infected cells. SIVmac1A11 and SIVmac1A11Δnef did not infect human tumor macrophage cell line U937, GCT, THP-1, or HL-60 cells, although virus was produced after these cells were transfected with either wild-type or nef mutant viral DNA. Similar levels of virus were recovered from U937 and THP-1 cells transfected with mutant and parental proviral DNAs. In transient expression assays in a T-cell line and a macrophage line, the nef protein of SIVmac1A11 did not significantly suppress or enhance expression of the chloramphenicol acetyltransferase reporter gene linked to the SIVmac long terminal repeat. Thus, abrogation of nef did not affect several in vitro properties of SIVmac1A11, including patterns of viral infection in rhesus PBMC, rhesus macrophages, or human T-cell lines.

Original languageEnglish (US)
Pages (from-to)5432-5442
Number of pages11
JournalJournal of Virology
Volume66
Issue number9
StatePublished - Sep 1992

Fingerprint

nef Genes
Simian immunodeficiency virus
Simian Immunodeficiency Virus
macrophages
Macrophages
Viruses
viruses
mononuclear leukocytes
virus replication
Blood Cells
cell lines
Macaca mulatta
genes
Cell Line
mutants
cells
T-lymphocytes
nef Gene Products
chloramphenicol acetyltransferase
T-Lymphocytes

ASJC Scopus subject areas

  • Immunology

Cite this

Unger, R. E., Marthas, M., Pratt-Lowe, E., Padrid, P. A., & Luciw, P. A. (1992). The nef gene of simian immunodeficiency virus SIVmac1A11. Journal of Virology, 66(9), 5432-5442.

The nef gene of simian immunodeficiency virus SIVmac1A11. / Unger, Ronald E.; Marthas, Marta; Pratt-Lowe, Elissa; Padrid, Phillip A.; Luciw, Paul A.

In: Journal of Virology, Vol. 66, No. 9, 09.1992, p. 5432-5442.

Research output: Contribution to journalArticle

Unger, RE, Marthas, M, Pratt-Lowe, E, Padrid, PA & Luciw, PA 1992, 'The nef gene of simian immunodeficiency virus SIVmac1A11', Journal of Virology, vol. 66, no. 9, pp. 5432-5442.
Unger RE, Marthas M, Pratt-Lowe E, Padrid PA, Luciw PA. The nef gene of simian immunodeficiency virus SIVmac1A11. Journal of Virology. 1992 Sep;66(9):5432-5442.
Unger, Ronald E. ; Marthas, Marta ; Pratt-Lowe, Elissa ; Padrid, Phillip A. ; Luciw, Paul A. / The nef gene of simian immunodeficiency virus SIVmac1A11. In: Journal of Virology. 1992 ; Vol. 66, No. 9. pp. 5432-5442.
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abstract = "The role of the simian immunodeficiency virus (SIV) nef gene in viral replication was investigated in several tissue culture systems. SIVmac1A11 is a molecularly cloned virus which replicates in both peripheral blood mononuclear cells (PBMC) and macrophages, although no disease is observed in infected rhesus macaques. In this report, we demonstrate that SIVmac1A11 contains a full open reading frame for nef which specifies a 37-kDa protein. To investigate the effects of nef on viral replication, a 70-bp deletion was introduced into the nef gene of SIVmac1A11. Analysis of infected cell extracts by immunoblotting revealed that both SIVmac1A11 and nef deletion virus SIVmac1A11Δnef produced the same viral proteins, except that Nef was absent in the mutant virus. The deletion mutation did not affect viral replication in PBMC, in monocyte-derived and alveolar macrophages obtained from rhesus macaques, and in human cell lines HUT-78 and CEMx-174. In addition, SIVmac1A11 and SIVmac1A11Δnef exhibited similar patterns of cytopathologic changes and ultrastructural appearances in infected cells. SIVmac1A11 and SIVmac1A11Δnef did not infect human tumor macrophage cell line U937, GCT, THP-1, or HL-60 cells, although virus was produced after these cells were transfected with either wild-type or nef mutant viral DNA. Similar levels of virus were recovered from U937 and THP-1 cells transfected with mutant and parental proviral DNAs. In transient expression assays in a T-cell line and a macrophage line, the nef protein of SIVmac1A11 did not significantly suppress or enhance expression of the chloramphenicol acetyltransferase reporter gene linked to the SIVmac long terminal repeat. Thus, abrogation of nef did not affect several in vitro properties of SIVmac1A11, including patterns of viral infection in rhesus PBMC, rhesus macrophages, or human T-cell lines.",
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