Analysis of rev gene function on human immunodeficiency virus type 1 replication in lymphoid cells by using a quantitative polymerase chain reaction method

S. J. Arrigo, S. Weitsman, Joseph D Rosenblatt, I. S Y Chen

Research output: Contribution to journalArticle

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Abstract

Most detailed anlayses of the human immunodeficiency virus type 1 (HIV-1) rev gene product have relied on transfection of subgenomic env constructs into cells in which amplification of the transfected DNA occurs. This was necessitated by difficulties in quantitating low-abundance HIV-1 mRNA species and in distinguishing different RNAs of similar sizes. We have modified the conventional polymerase chain reaction method for general use as an extremely sensitive procedure for quantitative analysis of RNA species. Using this method, we assessed the role of the HIV-1 rev gene in viral replication following mutagenesis of an infectious molecular clone, HIV-1(JR-CSF). Following transfection of wild-type and mutant proviral constructs, we can specifically detect unspliced RNA and distinguish between the spliced trat-rev and nef mRNAs, which are not resolved by standard RNA analyses. Our results show that the rev protein of HIV-1(JR-CSF) simultaneously down regulates the expression of tat-rev and nef RNAs and up regulates the level of unspliced full-length HIV-1 RNA. A cis-acting element(s), located exclusively within the env sequences, is essential to exhibit this regulation. Fractionation of cells shows that the ultimate effect of Rev is to direct the appearance of unspliced or singly spliced RNAs in the cytoplasm. Models are discussed for possible mechanisms of Rev action.

Original languageEnglish
Pages (from-to)4875-4881
Number of pages7
JournalJournal of Virology
Volume63
Issue number11
StatePublished - Jan 1 1989
Externally publishedYes

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rev Genes
Virus Replication
Human immunodeficiency virus 1
HIV-1
quantitative polymerase chain reaction
RNA
Lymphocytes
Polymerase Chain Reaction
genes
cells
transfection
Human Immunodeficiency Virus rev Gene Products
Transfection
methodology
rev Gene Products
cell fractionation
Cell Fractionation
Messenger RNA
virus replication
mutagenesis

ASJC Scopus subject areas

  • Immunology

Cite this

Analysis of rev gene function on human immunodeficiency virus type 1 replication in lymphoid cells by using a quantitative polymerase chain reaction method. / Arrigo, S. J.; Weitsman, S.; Rosenblatt, Joseph D; Chen, I. S Y.

In: Journal of Virology, Vol. 63, No. 11, 01.01.1989, p. 4875-4881.

Research output: Contribution to journalArticle

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