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PMID: 2839827 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Role of RNase H in hybrid-arrested translation by antisense oligonucleotides.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 14 ·1988-07-00 ·Pages 5011-5

Walder RY, Walder JA

Abstract

The mechanism of hybrid-arrested translation by antisense oligodeoxynucleotides has been investigated with the rabbit reticulocyte lysate system. The oligonucleotides studied were directed against different regions of mouse alpha- or beta-globin mRNAs. Freshly prepared reticulocyte lysates were found to contain 1-2% of the level of RNase H in nucleated cells. This level of activity was sufficient to cleave nearly 100% of the targeted mRNA at the site of hybridization with a complementary oligodeoxynucleotide in 1 hr under conditions of active translation. Using poly(rA).oligo(dT) as a competitive inhibitor of the enzyme, hybrid arrest by oligodeoxynucleotides complementary to the sequence spanning the initiation codon or to a sequence in the coding region was found to be due entirely to cleavage of mRNA by RNase H. Hybridization of oligodeoxynucleotides adjacent to the cap site of beta-globin mRNA, but not the alpha-globin mRNA, also inhibited protein synthesis directly. Even in this case, however, cleavage of the mRNA by RNase H was the predominant pathway of inhibition.

MeSH Terms
Animals Codon DNA/genetics Endoribonucleases/antagonists & inhibitors,metabolism Female Globins/genetics Mice Mice, Inbred BALB C Nucleic Acid Hybridization Oligodeoxyribonucleotides/pharmacology Oligonucleotides/genetics,pharmacology Oligonucleotides, Antisense Poly A/pharmacology Protein Biosynthesis/drug effects RNA, Messenger/genetics,metabolism Rabbits Reticulocytes/metabolism Ribonuclease H
Chemicals
Codon Oligodeoxyribonucleotides Oligonucleotides Oligonucleotides, Antisense RNA, Messenger Poly A poly(rA).oligo(dT) Globins DNA Endoribonucleases Ribonuclease H
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Walder R Y
Department of Biochemistry, University of Iowa, Iowa City 52242.
Walder J A
References (31)
31 references, click to expand
  1. Studies on a calf thymus ribonuclease specific for ribonucleic acid-deoxyribonucleic acid hybrids.
    Biochemistry. 1973 Jun 19;12(13):2389-95 PMID: 4709937
  2. Selective elimination of mRNAs in vivo: complementary oligodeoxynucleotides promote RNA degradation by an RNase H-like activity.
    Proc Natl Acad Sci U S A. 1987 Nov;84(22):7896-900 PMID: 2825169
  3. Interaction between the synthesis of alpha and beta globin.
    Eur J Biochem. 1975 Oct 15;58(2):339-50 PMID: 1183442
  4. An efficient mRNA-dependent translation system from reticulocyte lysates.
    Eur J Biochem. 1976 AUG 1;67(1):247-56 PMID: 823012
  5. Competition between globin messenger ribonucleic acids for a discriminating initiation factor.
    J Biol Chem. 1977 Apr 25;252(8):2684-90 PMID: 856800
  6. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange.
    Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835-8 PMID: 73185
  7. Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide.
    Proc Natl Acad Sci U S A. 1978 Jan;75(1):280-4 PMID: 75545
  8. Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide.
    Proc Natl Acad Sci U S A. 1978 Jan;75(1):285-8 PMID: 75546
  9. Resolution of hemoglobin subunits by electrophoresis in acid urea polyacrylamide gels containing Triton X-100.
    Anal Biochem. 1978 Apr;85(2):506-18 PMID: 646104
  10. Change of ribonuclease H activity from rat liver nuclei by thioacetamide: preferential increase in the activity of magnesium-dependent ribonuclease H.
    Biochem Biophys Res Commun. 1978 Nov 14;85(1):280-6 PMID: 743281
  11. DNA analysis in the diagnosis of hemoglobin disorders.
    Methods Enzymol. 1981;76:805-17 PMID: 6276651
  12. Translationally associated helix-destabilizing activity in rabbit reticulocyte lysate.
    J Biol Chem. 1984 Dec 25;259(24):15597-602 PMID: 6096379
  13. Constitutive and conditional suppression of exogenous and endogenous genes by anti-sense RNA.
    Science. 1985 Jul 26;229(4711):345-52 PMID: 2990048
  14. Quantitative hybridization-arrest of mRNA in Xenopus oocytes using single-stranded complementary DNA or oligonucleotide probes.
    Nucleic Acids Res. 1985 Jul 11;13(13):4991-5004 PMID: 3875079
  15. Inhibition of rabbit globin mRNA translation by sequence-specific oligodeoxyribonucleotides.
    Biochemistry. 1985 Oct 22;24(22):6132-8 PMID: 4084510
  16. Hybridization arrest of globin synthesis in rabbit reticulocyte lysates and cells by oligodeoxyribonucleoside methylphosphonates.
    Biochemistry. 1985 Oct 22;24(22):6139-45 PMID: 4084511
  17. Translation arrest by oligodeoxynucleotides complementary to mRNA coding sequences yields polypeptides of predetermined length.
    Nucleic Acids Res. 1986 Feb 11;14(3):1427-48 PMID: 3633502
  18. Red blood cells contain a pathway for the degradation of oxidant-damaged hemoglobin that does not require ATP or ubiquitin.
    J Biol Chem. 1986 May 5;261(13):5705-13 PMID: 3009430
  19. Antiviral effect of an oligo(nucleoside methylphosphonate) complementary to the splice junction of herpes simplex virus type 1 immediate early pre-mRNAs 4 and 5.
    Proc Natl Acad Sci U S A. 1986 May;83(9):2787-91 PMID: 3010316
  20. The 35-nucleotide spliced leader sequence is common to all trypanosome messenger RNA's.
    Science. 1986 Aug 1;233(4763):569-71 PMID: 3523758
  21. The common 5' terminal sequence on trypanosome mRNAs: a target for anti-messenger oligodeoxynucleotides.
    Nucleic Acids Res. 1986 Jul 25;14(14):5605-14 PMID: 3737413
  22. The role of antisense RNA in gene regulation.
    Annu Rev Biochem. 1986;55:569-97 PMID: 2427015
  23. The use of single-stranded DNA and RNase H to promote quantitative 'hybrid arrest of translation' of mRNA/DNA hybrids in reticulocyte lysate cell-free translations.
    Nucleic Acids Res. 1986 Aug 26;14(16):6433-51 PMID: 3018671
  24. Influence of 5' proximal secondary structure on the translational efficiency of eukaryotic mRNAs and on their interaction with initiation factors.
    J Biol Chem. 1986 Oct 25;261(30):13979-89 PMID: 3771516
  25. Anti-messenger oligodeoxynucleotides: specific inhibition of rabbit beta-globin synthesis in wheat germ extracts and Xenopus oocytes.
    Biochimie. 1986 Sep;68(9):1063-9 PMID: 3096384
  26. Inhibition of vesicular stomatitis virus protein synthesis and infection by sequence-specific oligodeoxyribonucleoside methylphosphonates.
    Biochemistry. 1986 Oct 7;25(20):6268-75 PMID: 3024704
  27. Specific antiviral activity of a poly(L-lysine)-conjugated oligodeoxyribonucleotide sequence complementary to vesicular stomatitis virus N protein mRNA initiation site.
    Proc Natl Acad Sci U S A. 1987 Feb;84(3):648-52 PMID: 3027696
  28. Specific hybridization arrest of dihydrofolate reductase mRNA in vitro using anti-sense RNA or anti-sense oligonucleotides.
    Arch Biochem Biophys. 1987 Feb 15;253(1):214-20 PMID: 2434026
  29. Antisense oligodeoxynucleotides provide insight into mechanism of translation initiation of two Sendai virus mRNAs.
    J Biol Chem. 1987 Jun 5;262(16):7492-6 PMID: 2438279
  30. Enzymatic amplification of translation inhibition of rabbit beta-globin mRNA mediated by anti-messenger oligodeoxynucleotides covalently linked to intercalating agents.
    Nucleic Acids Res. 1987 Jun 25;15(12):4717-36 PMID: 3037483
  31. Purification and properties of ribonuclease H of calf thymus.
    Proc Natl Acad Sci U S A. 1973 Jul;70(7):1959-63 PMID: 4516197
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.14.5011
Published
1988-07-00
Pages
5011-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC281677
Subset
IM
Grants
NIADDK NIH HHS · AM-25295 · United States
NHLBI NIH HHS · HL-33555 · United States
External Links
PubMed source DOI: 10.1073/pnas.85.14.5011 PMC Full Text View journal details
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