La Autoantigen Is Necessary for Optimal Function of the Poliovirus and Hepatitis C Virus Internal Ribosome Entry Site In Vivo and In Vitro

ABSTRACT Translation of poliovirus and hepatitis C virus (HCV) RNAs is initiated by recruitment of 40S ribosomes to an internal ribosome entry site (IRES) in the mRNA 5′ untranslated region. Translation initiation of these RNAs is stimulated by noncanonical initiation factors called IRES trans-activating factors (ITAFs). The La autoantigen is such an ITAF, but functional evidence for the role of La in poliovirus and HCV translation in vivo is lacking. Here, by two methods using small interfering RNA and a dominant-negative mutant of La, we demonstrate that depletion of La causes a dramatic reduction in poliovirus IRES function in vivo. We also show that 40S ribosomal subunit binding to HCV and poliovirus IRESs in vitro is inhibited by a dominant-negative form of La. These results provide strong evidence for a function of the La autoantigen in IRES-dependent translation and define the step of translation which is stimulated by La.

[1]  R. Korneluk,et al.  XIAP: Apoptotic brake and promising therapeutic target , 2001, Apoptosis.

[2]  N. Sonenberg,et al.  An efficient system for cap- and poly(A)-dependent translation in vitro. , 2004, Methods in molecular biology.

[3]  S. Tenenbaum,et al.  Differential phosphorylation and subcellular localization of La RNPs associated with precursor tRNAs and translation-related mRNAs. , 2003, Molecular cell.

[4]  Raul Andino,et al.  Nucleic Acid-Based Immune System: the Antiviral Potential of Mammalian RNA Silencing , 2003, Journal of Virology.

[5]  Mamoru Watanabe,et al.  Inhibition of intracellular hepatitis C virus replication by synthetic and vector‐derived small interfering RNAs , 2003, EMBO reports.

[6]  N. Sonenberg,et al.  Cell-Free Synthesis of Encephalomyocarditis Virus , 2003, Journal of Virology.

[7]  Saumitra Das,et al.  Hepatitis C Virus Internal Ribosome Entry Site-mediated Translation Is Stimulated by Specific Interaction of Independent Regions of Human La Autoantigen* , 2003, The Journal of Biological Chemistry.

[8]  Anastasia Khvorova,et al.  RNA interference blocks gene expression and RNA synthesis from hepatitis C replicons propagated in human liver cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[9]  F. Chisari,et al.  Interference of hepatitis C virus RNA replication by short interfering RNAs , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[10]  M. Caligiuri,et al.  BCR/ABL activates mdm2 mRNA translation via the La antigen. , 2003, Cancer cell.

[11]  Sophie Bonnal,et al.  IRESdb: the Internal Ribosome Entry Site database , 2003, Nucleic Acids Res..

[12]  C. Rice,et al.  Clearance of replicating hepatitis C virus replicon RNAs in cell culture by small interfering RNAs , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Bartenschlager.  Hepatitis C virus replicons: potential role for drug development , 2002, Nature Reviews Drug Discovery.

[14]  R. Andino,et al.  Short interfering RNA confers intracellular antiviral immunity in human cells , 2002, Nature.

[15]  G. Goodall,et al.  Hypoxia-inducible Factor-1 (cid:1) mRNA Contains an Internal Ribosome Entry Site That Allows Efficient Translation during Normoxia and Hypoxia , 2022 .

[16]  M. Zamora,et al.  Poliovirus-Mediated Shutoff of Host Translation: an Indirect Effect , 2002 .

[17]  R. Jackson.  Proteins Involved in the Function of Picornavirus Internal Ribosomal Entry Sites , 2002 .

[18]  B. Semler,et al.  Molecular biology of picornaviruses , 2002 .

[19]  T. Cedervall,et al.  The La protein. , 2002, Annual review of biochemistry.

[20]  Yoon Ki Kim,et al.  La autoantigen enhances translation of BiP mRNA. , 2001, Nucleic acids research.

[21]  Stéphan Vagner,et al.  Irresistible IRES , 2001, EMBO reports.

[22]  Nahum Sonenberg,et al.  Dual Interactions of the Translational Repressor Paip2 with Poly(A) Binding Protein , 2001, Molecular and Cellular Biology.

[23]  P. Sarnow,et al.  Internal ribosome entry sites in eukaryotic mRNA molecules. , 2001, Genes & development.

[24]  T. Tuschl,et al.  Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells , 2001, Nature.

[25]  T. Tuschl,et al.  RNA interference is mediated by 21- and 22-nucleotide RNAs. , 2001, Genes & development.

[26]  S. Schwartz,et al.  Binding of the La autoantigen to the hepatitis C virus 3' untranslated region protects the RNA from rapid degradation in vitro. , 2001, The Journal of general virology.

[27]  R. Cuesta,et al.  Preferential translation of adenovirus mRNAs in infected cells. , 2001, Cold Spring Harbor symposia on quantitative biology.

[28]  N. Sonenberg,et al.  Internal ribosome initiation of translation and the control of cell death. , 2000, Trends in genetics : TIG.

[29]  A. Siddiqui,et al.  Human La antigen is required for the hepatitis C virus internal ribosome entry site-mediated translation. , 2000, The Journal of biological chemistry.

[30]  T. Chow,et al.  Translational upregulation of X-linked inhibitor of apoptosis (XIAP) increases resistance to radiation induced cell death , 2000, Oncogene.

[31]  R. Korneluk,et al.  Functional Characterization of the X-Linked Inhibitor of Apoptosis (XIAP) Internal Ribosome Entry Site Element: Role of La Autoantigen in XIAP Translation , 2000, Molecular and Cellular Biology.

[32]  N. Sonenberg,et al.  Picornavirus RNA translation: roles for cellular proteins. , 2000, Trends in microbiology.

[33]  N. Sonenberg,et al.  A novel shuttling protein, 4E‐T, mediates the nuclear import of the mRNA 5′ cap‐binding protein, eIF4E , 2000, The EMBO journal.

[34]  S. Cornelis,et al.  Identification and characterization of a novel cell cycle-regulated internal ribosome entry site. , 2000, Molecular cell.

[35]  N. Sonenberg,et al.  A cell cycle-dependent internal ribosome entry site. , 2000, Molecular cell.

[36]  P. Sharp,et al.  RNAi Double-Stranded RNA Directs the ATP-Dependent Cleavage of mRNA at 21 to 23 Nucleotide Intervals , 2000, Cell.

[37]  Adi Kimchi,et al.  A Novel Form of DAP5 Protein Accumulates in Apoptotic Cells as a Result of Caspase Cleavage and Internal Ribosome Entry Site-Mediated Translation , 2000, Molecular and Cellular Biology.

[38]  R. Jackson.  4 A Comparative View of Initiation Site Selection Mechanisms , 2000 .

[39]  A. Hinnebusch.  5 Mechanism and Regulation of Initiator Methionyl-tRNA Binding to Ribosomes , 2000 .

[40]  Kenneth M. Kuhn,et al.  19 Cellular Internal Ribosome Entry Site Elements and the Use of cDNA Microarrays in Their Investigation , 2000 .

[41]  J. Hershey,et al.  2 The Pathway and Mechanism of Initiation of Protein Synthesis , 2000 .

[42]  N. Sonenberg,et al.  Translational control of gene expression , 2000 .

[43]  A. Nomoto,et al.  Lower concentration of La protein required for internal ribosome entry on hepatitis C virus RNA than on poliovirus RNA. , 1999, The Journal of general virology.

[44]  S. Kuge,et al.  Intracellular Redistribution of Truncated La Protein Produced by Poliovirus 3Cpro-Mediated Cleavage , 1999, Journal of Virology.

[45]  E. Wimmer,et al.  Poliovirus/Hepatitis C Virus (Internal Ribosomal Entry Site-Core) Chimeric Viruses: Improved Growth Properties through Modification of a Proteolytic Cleavage Site and Requirement for Core RNA Sequences but Not for Core-Related Polypeptides , 1999, Journal of Virology.

[46]  A. Gingras,et al.  eIF4 initiation factors: effectors of mRNA recruitment to ribosomes and regulators of translation. , 1999, Annual review of biochemistry.

[47]  R. Lloyd,et al.  Eukaryotic Translation Initiation Factor 4G Is Targeted for Proteolytic Cleavage by Caspase 3 during Inhibition of Translation in Apoptotic Cells , 1998, Molecular and Cellular Biology.

[48]  M. Ott,et al.  A Small Yeast RNA Blocks Hepatitis C Virus Internal Ribosome Entry Site (HCV IRES)-Mediated Translation and Inhibits Replication of a Chimeric Poliovirus under Translational Control of the HCV IRES Element , 1998, Journal of Virology.

[49]  A. Gingras,et al.  4E-BP3, a New Member of the Eukaryotic Initiation Factor 4E-binding Protein Family* , 1998, The Journal of Biological Chemistry.

[50]  S. Wolin,et al.  The Yeast La Protein Is Required for the 3′ Endonucleolytic Cleavage That Matures tRNA Precursors , 1997, Cell.

[51]  A. Siddiqui,et al.  The La antigen binds 5' noncoding region of the hepatitis C virus RNA in the context of the initiator AUG codon and stimulates internal ribosome entry site-mediated translation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[52]  J. Qin,et al.  Phosphorylation of the Human La Antigen on Serine 366 Can Regulate Recycling of RNA Polymerase III Transcription Complexes , 1997, Cell.

[53]  N. Sonenberg,et al.  translation . domain that is essential for enhancing The La autoantigen contains a dimerization , 1996 .

[54]  G. Dreyfuss,et al.  General RNA binding proteins render translation cap dependent. , 1996, The EMBO journal.

[55]  G. Pruijn,et al.  Interaction of the La (SS-B) autoantigen with small ribosomal subunits. , 1996, European journal of biochemistry.

[56]  E. Wimmer,et al.  Poliovirus chimeras replicating under the translational control of genetic elements of hepatitis C virus reveal unusual properties of the internal ribosomal entry site of hepatitis C virus. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[57]  R. Jackson,et al.  Unique features of internal initiation of hepatitis C virus RNA translation. , 1995, The EMBO journal.

[58]  R. Jackson,et al.  Internal initiation of translation in eukaryotes: the picornavirus paradigm and beyond. , 1995, RNA.

[59]  N. Sonenberg,et al.  Repression of cap‐dependent translation by 4E‐binding protein 1: competition with p220 for binding to eukaryotic initiation factor‐4E. , 1995, The EMBO journal.

[60]  J. Flanegan,et al.  Complete replication of poliovirus in vitro: preinitiation RNA replication complexes require soluble cellular factors for the synthesis of VPg-linked RNA , 1995, Journal of virology.

[61]  N. Sonenberg,et al.  Translational control of gene expression , 2000 .

[62]  N. Gray,et al.  Iron regulatory protein prevents binding of the 43S translation pre‐initiation complex to ferritin and eALAS mRNAs. , 1994, The EMBO journal.

[63]  V. Agol,et al.  Internal translation initiation on poliovirus RNA: further characterization of La function in poliovirus translation in vitro , 1994, Journal of virology.

[64]  N. Sonenberg,et al.  Internal initiation of picornavirus RNA translation , 1993 .

[65]  V. Agol,et al.  La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate , 1993, Journal of virology.

[66]  V. Agol,et al.  Prokaryotic-like cis elements in the cap-independent internal initiation of translation on picornavirus RNA , 1992, Cell.

[67]  A. Paul,et al.  Cell-free, de novo synthesis of poliovirus. , 1991, Science.

[68]  N. Sonenberg,et al.  In vitro mutational analysis of cis-acting RNA translational elements within the poliovirus type 2 5' untranslated region , 1991, Journal of virology.

[69]  S. Le,et al.  Structural and functional analysis of the ribosome landing pad of poliovirus type 2: in vivo translation studies , 1991, Journal of virology.

[70]  N. Sonenberg,et al.  A cellular protein that binds to the 5'-noncoding region of poliovirus RNA: implications for internal translation initiation. , 1989, Genes & development.

[71]  N. Sonenberg,et al.  Mutational analysis of upstream AUG codons of poliovirus RNA , 1988, Journal of virology.

[72]  E. Wimmer,et al.  A segment of the 5' nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation , 1988, Journal of virology.

[73]  N. Sonenberg,et al.  Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA , 1988, Nature.

[74]  V. Agol,et al.  The genomes of attenuated and virulent poliovirus strains differ in their in vitro translation efficiencies. , 1985, Virology.

[75]  B. Semler,et al.  In vitro translation of poliovirus RNA: utilization of internal initiation sites in reticulocyte lysate , 1984, Journal of virology.

[76]  M. Pallansch,et al.  [46] Preparation and characterization of encephalornyocarditis (EMC) virus , 1981 .

[77]  M. Pallansch,et al.  Preparation and characterization of encephalomyocarditis (EMC) virus. , 1981, Methods in enzymology.

[78]  B. Semler,et al.  Protein-linked RNA of poliovirus is competent to form an initiation complex of translation in vitro , 1980, Nature.

[79]  E. Ehrenfeld,et al.  Translation of poliovirus RNA in vitro: changes in cleavage pattern and initiation sites by ribosomal salt wash. , 1979, Virology.

[80]  N. Jones,et al.  Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells , 1979, Cell.