Selective inhibition of internally initiated RNA translation

Details Selective inhibition of internally initiated RNA translation Selective inhibition of internally initiated RNA translation

1997-10-06

1999-11-23

LeGuyader, John

Chemistry: molecular biology and microbiology

Vector, per se

435 6, 435325, 435375, 435455, 536 241, 536 245, C07H 2102, C07H 2104, C12N 1585

Patent

active

059899042

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

The invention relates to selective inhibition of translation of certain mRNAs. More particularly, the invention relates to selective inhibition of an mRNA which is initiated at an internal ribosome entry site, such as a picornavirus RNA, by a small RNA or a molecular mimic thereof. This RNA or mimic interacts specifically with a cellular protein to prevent binding of that protein to the internal ribosome entry site, thereby inhibiting translation initiation at that entry site.


BACKGROUND ART

Picornaviruses include inter alia polioviruses, which cause infantile paralysis, and rhinoviruses, which cause the common cold. Picorna-related viruses, which replicate by mechanisms similar to picornaviruses, include hepatitis A and C, major causes of human hepatitis. Although poliovirus vaccines are available, cases of polio still develop where vaccination is not properly used. Vaccines for other picornaviruses may not be feasible, for instance, due to the high rate of mutability of the viral coat proteins in the rhinoviruses. Therefore, there is a need for methods and compositions for selectively inhibiting picornavirus replication without toxic effects on the host cells.
Poliovirus, the prototype member of the picornaviridae family, is a single stranded, plus-sense RNA virus which multiplies in the cytoplasm of infected cells. The RNA genome comprises approximately 7,500 nucleotides and codes for a 250 kDa polyprotein (Kitamura, N. et al. Nature (1981) 291:547-553 and Racaniello, V. R., et al. Proc Natl Acad Sci USA (1981) 78:4887-4891. The unusually long 5' untranslated region (5'UTR) of poliovirus RNA (750 nucleotides) is highly structured (Skinner, M. A. et al. J Mol Biol (1989) 207:379-392; Agol, V. Adv Virus Res (1991) 40:103-180) and contains six to eight upstream AUGs, none of which appears to be used in initiation of translation (Pelletier, J. et al. J Virol (1988a) 62:4486-4492.
Translation of most mammalian cellular mRNAs proceeds by binding of ribosomes to the 5' cap structure followed by scanning of the mRNA until the appropriate AUG is encountered by the ribosome (Kozak, M. Microbiol Rev (1983) 47:1-45). In contrast translation of naturally uncapped poliovirus RNA has been shown to be mediated by a mechanism involving internal entry of ribosomes near the initiator AUG (Pelletier, J. et al. Nature (1988) 334:320-325). Recent studies have demonstrated that internal entry of ribosomes requires an element located between nucleotides 320-631 within the 5'UTR of poliovirus RNA (Pelletier, J. et al., supra). This sequence element has been termed a ribosome landing pad (RLP) or, more generally, internal ribosome entry site (IRES). Although a number of cellular polypeptides have been implicated in IRES-dependent translation, the precise mechanism of internal initiation of translation remains poorly understood.
In addition to poliovirus many other picornaviruses have been shown to utilize this novel mechanism for initiation of translation (Jang, S. K. et al. Genes Dev (1990) 4:1560-1572, Belsham, G. J. et al. J Virol (1990) 64:5389-5395, Jackson, R. et al. Trends Biochem Sci (1990) 15:477-483, Luz, N. et al. FEBS Letters (1990) 269:311-314, Luz, N. et al. Virology (1991) 65:6486-6494, Bandopadhyay, P. K. et al. J Virol (1992) 66:6249-6256, Borman, A. et al. Virology (1992) 188:685-696, Borman, A. et al. Gen Virol (1993) 74:1775-1788). The RNA genomes of two picorna-related viruses, hepatitis A and C, have been shown to utilize internal ribosome entry for translation initiation (Kohara, K. T. et al. J Virol (1992) 66:1476-1483 and Glass, M. J. et al. Virology (1993) 193:842-852). Two cellular mRNAs, encoding immunoglobulin heavy chain binding protein (Bip), the mouse androgen receptor (32) and the antennapedia of Drosophila, also have been shown to use internal initiation of translation (Macejak, D. G. et al. Nature (1991) 353:90-94 and Oh, S. K. et al. Genes Dev (1992) 6:1643-1653).
All picornaviral mRNAs that utilize IRES-dependent translation contain a polypyrimidine tract located at the 3'

REFERENCES:
Agrawal, S. "Antisense Oligonucleotides: Towards Clinical Trials" TIBTECH vol. 14:376-387, 1996.
Stull et al "Antigene, Ribozyme and Aptamer Nucleic Acid Drugs: Progress and Prospects" Pharmaceutical Research vol. 12(4):465-483, 1995.
Kitamura, N. et al. Primary Structure, Gene Organization and Polypeptide Expression of Poliovirus RNA, Nature (1981) 291:547-553.
Racaniello, V.R., et al. Molecular Cloning of Poliovirus cDNA and Determination of the Complete Nucleotide Sequence of the Viral Genome, Proc Natl Acad Sci USA (1981) 78:4887-4891.
Skinner, M.A. et al. New Model for the Secondary Structure of the 5' Non-coding RNA of Poliovirus is Supported by Biochemical and Genetic Data Tha Also Show That RNA Secondary Structure is Important in Neurovirulence, J Mol Biol (1989) 207:379-392.
Agol, V., The 5'-Untranslated Region of Picornaviral Genomes, Adv Virus Res (1991) 40:103-180.
Pelletier, J. et al. Mutational Analysis of Upstream AUG Codons of Polivirus RNA, J. Virol (1988a) 62:4486-4492.
Kozak, M., Comparison of Initiation of Protein Synthesis in Procaryotes, Eucaryotes, and Organelles, Microbiol Rev (1983) 47:1-45.
Pelletier, J. et al. Internal Initiation of Translation of Eukaryotic mRNA Directed by a Sequence Derived from Poliovirus RNA, Nature (1988) 334:320-325.
Jang, S.K. et al. Cap-independent Translation of Encephalomyocarditis Virus RNA: Structural Elements of the Internal Ribonsomal Entry Site and Involvement of a Cellular 57-kD RNA-binding Protein, Genes Dev (1990) 4:1560-1572.
Belsham, G.J. et al. A Region of the 5' Noncoding Region of Foot-and-Mouth Disease Virus RNA Directs Efficient Internal Initiation of Protein Synthesis Within Cells: Involvement with the Role of L Protease in Translational Control, J Virol (1990) 64:5389-5395.
Jackson, R. et al. The Novel Mechanism of Initiation of Picornavirus RNA Translation, Trends Biochem Sci (1990) 15:477-483.
Luz, N. et al. A Cellular 57 kDa Protein Binds to Two Regions of the Internal Translation Initiation Site of Foot-and-Mouth Disease Virus, FEBS Letters (1990) 269:311-314.
Luz, N. et al. Interaction of a Cellular 57-Kilodalton Protein with the Internal Translation Initiation Site of Foot-and-Mouth Disease Virus Virology (1991) 65:6486-6494.
Bandopadhyay, P.K. et al. Cap-Independent Translation by the 5' Untranslated Region of Theiler's Murine Encephalomyelitis Virus, J Virol (1992) 66:6249-6256.
Borman, A. et al. Initiation of Translation of Human Rhinovirus RNA: Mapping the Internal Ribosome Entry Site, Virology (1992) 188:685-696.
Borman, A. et al. The Involvement of a Spliceosome Component in Internal Initiation of Human Rhinovirus RNA Translation, Gen Virol (1993) 74:1775-1788.
Kohara, K.T. et al. Internal Ribosome Entry Site within Hepatitis C Virus RNA, J Virol (1992) 66:1476-1483.
Glass, M.J. et al. Identification of the Hepatitis A Virus Internal Ribosome Entry Site: In Vivo and In Vitro Analysis of Bicistronic RNAs Containing the HAV 5' Noncoding Region, Virology (1993) 193:842-852.
Macejak, D.G. et al. Internal Initiation of Translation Mediated by the 5' Leader of a Cellular mRNA, Nature (1991) 353:90-94.
Oh, S.K. et al. Homeotic Gene Antennapaedia mRNA Contains 5'-Noncoding Sequences that Confer Translational Initiation by Internal Ribonsome Binding, Genes Dev (1992) 6:1643-1653.
Pilipenko, E.V. et al. Prokaryotic-like Cis Elements in the Cap-Independent Internal Initiation of Translation on Picornavirus RNA, Cell (1992) 68:119-131.
Brown, B.A. et al. Translation of Poliovirus RNA In Vitro: Changes in Cleavage Pattern and Initiation Sites by Ribonsomal Salt Wash, Virology (1979) 97:376-405.
Dorner, H.A. et al. In Vitro Translation of Poliovirus RNA: Utilization of Internal Initiation Sites in Reticulocyte Lysate, J. Virol (1984) 50:507-514.
Najita, L., Oxidation--Reduction Sensitive Interaction of a Cellular 50-kDa Protein with an RNA Hairpin in the 5' Noncoding Region of the Poliovirus Genome, Proc Natl Acad Sci USA (1990) 87:5846-5850.
Meerovitch, K. et al., A Cellular

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