1887

Abstract

The unique poly(U/UC) tract, the middle part of the tripartite 3′ non-coding region (3′NCR) of hepatitis C virus (HCV) genomic RNA, may represent a recognition signal for the HCV replicase complex. In this study, several proteins binding specifically to immobilized ribooligonucleotide r(U) mimicking this structure were identified using cytosolic extracts from HCV-negative or -positive liver explants, and a prominent 36 kDa protein was studied further. Competition experiments including homoribopolymers revealed binding affinities in the order: oligo/poly(U)≫(A)≫(C)≫(G). The protein was identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a multifunctional protein known to bind RNA. GAPDH bound efficiently to the full-length HCV RNA and binding to various 3′NCR constructs revealed critical dependence upon the presence of the middle part of the 3′NCR. Polypyrimidine tract-binding protein, described previously to bind the 3′NCR, did not bind efficiently to the middle part of 3′NCR and was captured from liver extracts in considerably smaller quantities.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-80-12-3109
1999-12-01
2024-10-08
Loading full text...

Full text loading...

/deliver/fulltext/jgv/80/12/0803109.html?itemId=/content/journal/jgv/10.1099/0022-1317-80-12-3109&mimeType=html&fmt=ahah

References

  1. Behrens S.-E., Tomei L., De Francesco R. 1996; Identification and properties of the RNA-dependent RNA polymerase of hepatitis C virus. EMBO Journal 15:12–22
    [Google Scholar]
  2. Chung R. T., Kaplan L. M. 1999; Heterogeneous nuclear ribonucleoprotein I (hnRNP-I/PTB) selectively binds to the conserved 3′ terminus of hepatitis C viral RNA. Biochemical and Biophysical Research Communications 254:351–362
    [Google Scholar]
  3. Clarke B. 1997; Molecular virology of hepatitis C virus. Journal of General Virology 78:2397–2410
    [Google Scholar]
  4. De B. P., Gupta S., Zhao H., Drazba J. A., Banerjee A. K. 1996; Specific interaction in vitro and in vivo of glyceraldehyde-3-phosphate dehydrogenase and LA protein with cis-acting RNAs of human parainfluenza virus type 3. Journal of Biological Chemistry 271:24728–24735
    [Google Scholar]
  5. Dreyfuss G., Matunis M. J., Pinol-Roma S., Burd C. G. 1993; hnRNP proteins and the biogenesis of mRNA. Annual Review of Biochemistry 62:289–321
    [Google Scholar]
  6. Furuya T., Lai M. M. C. 1993; Three different cellular proteins bind to complementary sites on the 5′-end-positive and 3′-end-negative strands of mouse hepatitis virus RNA. Journal of Virology 67:7215–7222
    [Google Scholar]
  7. Glasser P. E., Gross R. W. 1995; Rapid plasmenylethanolamine-selective fusion of membrane bilayers catalyzed by an isoform of glyceraldehyde-3-phosphate dehydrogenase: discrimination between glycolytic and fusogenic roles of individual isoforms. Biochemistry 34:12193–12203
    [Google Scholar]
  8. Gontarek R. R., Gutshall L. L., Herold K. M., Tsai J., Sathe G. M., Mao J., Prescott C., Del Vecchio A. M. 1999; hnRNP C and polypyrimidine tract-binding protein specifically interact with the pyrimidine-rich region within the 3′NTR of the HCV RNA genome. Nucleic Acids Research 27:1457–1463
    [Google Scholar]
  9. Huitorel P., Pantaloni D. 1985; Bundling of microtubules by glyceraldehyde-3-phosphate dehydrogenase and its modulation by ATP. European. Journal of Biochemistry 150:265–269
    [Google Scholar]
  10. Hunt S. L., Jackson R. J. 1999; Polypyrimidine-tract binding protein (PTB) is necessary, but not sufficient, for efficient internal initiation of translation of human rhinovirus-2 RNA. RNA 5:344–359
    [Google Scholar]
  11. Ito T., Lai M. M. C. 1997; Determination of the secondary structure of and cellular protein binding to the 3′-untranslated region of the hepatitis C virus RNA genome. Journal of Virology 71:8698–8706
    [Google Scholar]
  12. Ito T., Lai M. M. C. 1999; An internal polypyrimidine-tract-binding protein-binding site in the hepatitis C virus RNA attenuates translation, which is relieved by the 3′-untranslated sequence. Virology 254:288–296
    [Google Scholar]
  13. Kanai A., Tanabe K., Kohara M. 1995; Poly(U) binding activity of hepatitis C virus NS3 protein, a putative RNA helicase. FEBS Letters 376:221–224
    [Google Scholar]
  14. Karpel R. L., Burchard A. C. 1981; A basic isozyme of yeast glyceraldehyde-3-phosphate dehydrogenase with nucleic acid helix-destabilizing activity. Biochimica et Biophysica Acta 654:256–267
    [Google Scholar]
  15. Kolykhalov A. A., Feinstone S. M., Rice C. M. 1996; Identification of a highly conserved sequence element at the 3′ terminus of hepatitis C virus genome RNA. Journal of Virology 70:3363–3371
    [Google Scholar]
  16. Kolykhalov A. A., Agapov E. V., Blight K. J., Mihalik K., Feinstone S. M., Rice C. M. 1997; Transmission of hepatitis C by intrahepatic inoculation with transcribed RNA. Science 277:570–574
    [Google Scholar]
  17. Kusov Y., Weitz M., Dollenmeier G., Gauss-Muller V., Siegl G. 1996; RNA–protein interactions at the 3′ end of the hepatitis A virus RNA. Journal of Virology 70:1890–1897
    [Google Scholar]
  18. Luo G. 1999; Cellular proteins bind to the poly(U) tract of the 3′ untranslated region of hepatitis C virus RNA genome. Virology 256:105–118
    [Google Scholar]
  19. Major M. E., Feinstone S. M. 1997; The molecular virology of hepatitis C. Hepatology 25:1527–1538
    [Google Scholar]
  20. Mejean C., Pons F., Benyamin Y., Roustan C. 1989; Antigenic probes locate binding sites for the glycolytic enzymes glyceraldehyde-3-phosphate dehydrogenase, aldolase and phosphofructokinase in the actin monomer in microfilaments. Biochemical Journal 264:671–677
    [Google Scholar]
  21. Morgenstern K. A., Landro J. A., Hsiao K., Lin C., Gu Y., Su M. S.-S., Thomson J. A. 1997; Polynucleotide modulation of the protease, nucleoside triphosphatase, and helicase activities of a hepatitis C virus NS3–NS4A complex isolated from transfected COS cells. Journal of Virology 71:3767–3775
    [Google Scholar]
  22. Nagy E., Rigby W. F. C. 1995; Glyceraldehyde-3-phosphate dehydrogenase selectively binds AU-rich RNA in the NAD+-binding region (Rossmann fold. Journal of Biological Chemistry 270:2755–2763
    [Google Scholar]
  23. Petrik J., Pearson G. M. J., Allain J.-P. 1997; High throughput PCR detection of HCV based on semiautomated multisample RNA capture. Journal of Virological Methods 64:147–159
    [Google Scholar]
  24. Petrik J., Parker H., Alexander G. J. M. 1998; Interactions of hepatitis C virus RNA non-coding regions with liver cell proteins. Hepatology 28:363A (Abstract
    [Google Scholar]
  25. Saunders P. A., Chen R. W., Chuang D. M. 1999; Nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase isoforms during neuronal apoptosis. Journal of Neurochemistry 72:925–932
    [Google Scholar]
  26. Schultz D. E., Hardin C. C., Lemon S. M. 1996; Specific interaction of glyceraldehyde 3-phosphate dehydrogenase with the 5′-nontranslated RNA of hepatitis A virus. Journal of Biological Chemistry 271:14134–14142
    [Google Scholar]
  27. Sella O., Gerlitz G., Le S. Y., Elroy-Stein O. 1999; Differentiation-induced internal translation of c-sis mRNA: analysis of the cis elements and their differentiation-linked binding to the hnRNP C protein. Molecular and Cellular Biology 19:5429–5440
    [Google Scholar]
  28. Singh R., Green M. R. 1993; Sequence-specific binding of transfer RNA by glyceraldehyde-3-phosphate dehydrogenase. Science 259:365–368
    [Google Scholar]
  29. Tanaka T., Kato N., Cho M.-J., Shimotohno K. 1995; A novel sequence found at the 3′ terminus of hepatitis C virus genome. Biochemical and Biophysical Research Communications 215:744–749
    [Google Scholar]
  30. Todd S., Nguyen J. H. C., Semler B. L. 1995; RNA–protein interactions directed by the 3′ end of human rhinovirus genomic RNA. Journal of Virology 69:3605–3614
    [Google Scholar]
  31. Tsuchihara K., Tanaka T., Hijikata M., Kuge S., Toyoda H., Nomoto A., Yamamoto N., Shimotohno K. 1997; Specific interaction of polypyrimidine tract-binding protein with the extreme 3′-terminal structure of hepatitis C virus genome, the 3′X. Journal of Virology 71:6720–6726
    [Google Scholar]
  32. Yamada N., Tanihara K., Takada A., Yorihuzi T., Tsutsumi M., Shimomura H., Tsuji T., Date T. 1996; Genetic organization and diversity of the 3′ noncoding region of the hepatitis C virus genome. Virology 223:255–261
    [Google Scholar]
  33. Yanagi M., St Claire M., Emerson S. U., Purcell R. H., Bukh J. 1999; In vivo analysis of the 3′ untranslated region of the hepatitis C virus after in vitro mutagenesis of an infectious cDNA clone. Proceedings of the National Academy of Sciences, USA 96:2291–2295
    [Google Scholar]
  34. Yu W., Leibowitz J. L. 1995; Specific binding of host cellular proteins to multiple sites within the 3′ end of mouse hepatitis virus genomic RNA. Journal of Virology 69:2016–2023
    [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-80-12-3109
Loading
/content/journal/jgv/10.1099/0022-1317-80-12-3109
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error