Identification of a sequence element immediately upstream of the polypurine tract that is essential for replication of simian immunodeficiency virus

P. O. Ilyinskii, Ronald Charles Desrosiers

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

A short stretch of T-rich sequences immediately upstream of the polypurine tract (PPT) is highly conserved in the proviral genomes of human and simian immunodeficiency viruses (HIV and SIV). To investigate whether this 'U-box' influences SIVmac239 replication, we analyzed the properties of mutants with changes in this region of the viral genome. All mutants were either retarded in their growth (up to one month delay) or did not replicate detectably in CEMx174 cells. When U-box mutants did replicate detectably, compensatory changes were consistently observed in the viral genome. The most common compensatory change was the acquisition of thymidines immediately upstream of the PPT, but marked expansion in the length of the PPT was also observed. U-box mutants produced transiently by transfection were severely impaired in their ability to produce reverse transcripts in infectivity assays. Analysis of transiently produced mutant virus revealed no defect in RNA packaging or virus assembly. These results identify a new structural element important for an early step in the viral life cycle that includes reverse transcription.

Original languageEnglish (US)
Pages (from-to)3766-3774
Number of pages9
JournalEMBO Journal
Volume17
Issue number13
DOIs
StatePublished - Jul 1 1998
Externally publishedYes

Fingerprint

Simian Immunodeficiency Virus
Viral Genome
Viruses
Genes
HIV
Virus Assembly
Product Packaging
Life Cycle Stages
Thymidine
Reverse Transcription
Transfection
Genome
RNA
Transcription
Life cycle
Assays
Packaging
Growth
Defects

Keywords

  • HIV
  • Replication
  • Reverse transcription
  • SIV
  • U-box

ASJC Scopus subject areas

  • Cell Biology
  • Genetics

Cite this

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N2 - A short stretch of T-rich sequences immediately upstream of the polypurine tract (PPT) is highly conserved in the proviral genomes of human and simian immunodeficiency viruses (HIV and SIV). To investigate whether this 'U-box' influences SIVmac239 replication, we analyzed the properties of mutants with changes in this region of the viral genome. All mutants were either retarded in their growth (up to one month delay) or did not replicate detectably in CEMx174 cells. When U-box mutants did replicate detectably, compensatory changes were consistently observed in the viral genome. The most common compensatory change was the acquisition of thymidines immediately upstream of the PPT, but marked expansion in the length of the PPT was also observed. U-box mutants produced transiently by transfection were severely impaired in their ability to produce reverse transcripts in infectivity assays. Analysis of transiently produced mutant virus revealed no defect in RNA packaging or virus assembly. These results identify a new structural element important for an early step in the viral life cycle that includes reverse transcription.

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