Sequence-specific interaction between HIV-1 matrix protein and viral genomic RNA revealed by in vitro genetic selection

Prakash Purohit, Stefan Dupont, Mario Stevenson, Michael R. Green

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

The human immunodeficiency virus type-1 matrix protein (HIV-1 MA) is a multifunctional structural protein sized as part of the Pr55 gag polyprotein. We have used in vitro genetic selection to identify an RNA consensus sequence that specifically interacts with MA (Kd = 5 × 10-7 M). This 13-nt MA binding consensus sequence bears a high degree of homology (77%) to a region (nt 1433-1446) within the POL open reading frame of the HIV-1 genome (consensus sequence from 38 HIV-1 strains). Chemical interference experiments identified the nucleotides within the MA binding consensus sequence involved in direct contact with MA. We further demonstrate that this RNA-protein interaction is mediated through a stretch of basic amino acids within MA. Mutations that disrupt the interaction between MA and its RNA binding site within the HIV-1 genome resulted in a measurable decrease in viral replication.

Original languageEnglish
Pages (from-to)576-584
Number of pages9
JournalRNA
Volume7
Issue number4
DOIs
StatePublished - May 20 2001
Externally publishedYes

Fingerprint

Genetic Selection
Consensus Sequence
Viral RNA
HIV-1
Proteins
Genome
RNA
gag Gene Products
Basic Amino Acids
Open Reading Frames
Nucleotides
Binding Sites
Mutation
In Vitro Techniques

Keywords

  • HIV
  • In vitro selection
  • Matrix protein
  • RNA-protein interaction

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Cite this

Sequence-specific interaction between HIV-1 matrix protein and viral genomic RNA revealed by in vitro genetic selection. / Purohit, Prakash; Dupont, Stefan; Stevenson, Mario; Green, Michael R.

In: RNA, Vol. 7, No. 4, 20.05.2001, p. 576-584.

Research output: Contribution to journalArticle

Purohit, Prakash ; Dupont, Stefan ; Stevenson, Mario ; Green, Michael R. / Sequence-specific interaction between HIV-1 matrix protein and viral genomic RNA revealed by in vitro genetic selection. In: RNA. 2001 ; Vol. 7, No. 4. pp. 576-584.
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