Control of ribonucleic acid function by oligonucleoside methylphosphonates

Paul S. Miller, Cheryl H. Agris, Laure Aurelian, Kathleen R. Blake, Akira Murakami, M. Parameswara Reddy, Sharon A. Spitz, Paul O.P. Ts'O

Research output: Contribution to journalArticlepeer-review

Abstract

Oligodeoxyribonucleoside methylphosphonates contain nonionic 3′-5′ linked methylphosphonate internucleotide bonds in place of the normal charged phosphodiester linkage of natural nucleic acids. These oligomers are resistant to nuclease hydrolysis, can pass through the membranes of mammalian cells in culture and can form stable hydrogen-bonded complexes with complementary nucleotide sequences of cellular RNAs such as mRNA. The oligomers are readily synthesized on insoluble polymer supports. Their chainlength and nucleotide sequence can be determined by chemical sequencing procedures. Oligonucleoside methylphosphonates which are complementary to the 5′-end, initiation codon region, or coding region of rabbit globin mRNA inhibit translation of the mRNA in rabbit reticulocyte lysates and globin synthesis in rabbit reticulocytes. This inhibition is due to the interaction of the oligomers with mRNA and the extent of inhibition is influenced by the secondary structure of the mRNA and the location of oligomer binding site on the mRNA. Oligomers complementary to the initiation codon regions of N, NS and G protein mRNAs of Vesicular stomatitis virus (VSV) inhibit virus protein synthesis in VSV-infected Mouse l-cells. These oligomers do not affect l-cell protein synthesis or growth. Virus protein synthesis and growth can also be selectively inhibited by oligonucleoside methylphosphonates which are complementary to the donor or acceptor splice junctions of virus pre mRNA. An oligomer complementary to the donor splice junction of SV40 large T antigen mRNA inhibits T-antigen synthesis in SV40-infected African green monkey kidney cells but does not inhibit overall cellular protein synthesis. An oligomer complementary to the acceptor splice junctions of Herpes simplex virus type 1 (HSV-1) immediate early mRNA 4 and 5 reduces virus production by two log units in HSV-1 and HSV-2 infected Vero cells. This oligomer does not affect the viability of the Vero cells. The results of these studies suggest that oligodeoxyribonucleoside methylphosphonates may be used to specifically control the expression of genes in living cells by interfering with the translation or splicing of mRNA. Because the specificity of these compounds resides in their ability to bind to complementary nucleic acid sequences, it should be possible to design oligomers which can be used to probe and perhaps control the function of specific proteins in normal, transformed, or virus-infected cells.

Original languageEnglish (US)
Pages (from-to)769-776
Number of pages8
JournalBiochimie
Volume67
Issue number7-8
DOIs
StatePublished - 1985

Keywords

  • hybridization arrest
  • messenger RNA
  • oligodeoxyribonucleoside methylphosphonates
  • splicing
  • translation

ASJC Scopus subject areas

  • Biochemistry

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