Previous Article | Next Article 
Journal of Virology, February 2001, p. 1348-1358, Vol. 75, No. 3
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.3.1348-1358.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Hepatitis C Virus 3'X Region Interacts with
Human Ribosomal Proteins
Jonny
Wood,1
Robert M.
Frederickson,2,
Stanley
Fields,2 and
Arvind H.
Patel1,*
MRC Virology Unit, Institute of Virology,
Glasgow G11 5JR, United Kingdom,1 and
Howard Hughes Medical Institute, Departments of Genetics and
Medicine, University of Washington, Seattle, Washington
98195-73602
Received 14 August 2000/Accepted 7 November 2000
To identify proteins that can bind the 3' untranslated region (UTR)
of hepatitis C virus (HCV) we screened human cDNA libraries using the
Saccharomyces cerevisiae three-hybrid system. Screening with an RNA sequence derived from the 3'-terminal 98 nucleotides (3'X
region) of an infectious clone of HCV (H77c) yielded clones of human
ribosomal proteins L22, L3, S3, and mL3, a mitochondrial homologue of
L3. We performed preliminary characterization of the binding between
the 3'X region and these proteins by a three-hybrid mating assay using
mutant 3'X sequences. We have further characterized the interaction
between 3'X and L22, since this protein is known to be associated with
two small Epstein-Barr virus (EBV)-encoded RNA species (EBERs) which
are abundantly produced in cells latently infected with EBV. The EBERs,
which have similar predicted secondary structure to the HCV 3'X,
assemble into ribonucleoprotein particles that include L22 and La
protein. To confirm that L22 binds HCV 3'X we performed in vitro
binding assays using recombinant L22 (expressed as a glutathione
S-transferase [GST] fusion protein) together with a 3'X
riboprobe. The 3'X region binds to the GST-L22 fusion protein (but not
to GST alone), and this interaction is subject to competition with
unlabeled 3'X RNA. To establish the functional role played by L22 in
internal ribosome entry site (IRES)-mediated translation of HCV
sequences we performed translational analysis in HuH-7 cells using
monocistronic and bicistronic reporter constructs. The relative amount
of core-chloramphenicol acetyltransferase reporter protein translated
under the control of the HCV IRES was stimulated in the presence of L22
and La when these proteins were supplied in trans.
*
Corresponding author. Mailing address: MRC Virology
Unit, Institute of Virology, Church St., Glasgow G11 5JR, United
Kingdom. Phone: 44 141 330 4026. Fax: 44 141 337 2236. E-mail:
a.patel{at}vir.gla.ac.uk.

Present address: Nature America, Inc., New York, NY
10010-1707.
Journal of Virology, February 2001, p. 1348-1358, Vol. 75, No. 3
0022-538X/01/$04.00+0 DOI: 10.1128/JVI.75.3.1348-1358.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Houmani, J. L., Davis, C. I., Ruf, I. K.
(2009). Growth-Promoting Properties of Epstein-Barr Virus EBER-1 RNA Correlate with Ribosomal Protein L22 Binding. J. Virol.
83: 9844-9853
[Abstract]
[Full Text]
-
Romero-Lopez, C., Berzal-Herranz, A.
(2009). A long-range RNA-RNA interaction between the 5' and 3' ends of the HCV genome. RNA
15: 1740-1752
[Abstract]
[Full Text]
-
Murakami, K., Kimura, T., Osaki, M., Ishii, K., Miyamura, T., Suzuki, T., Wakita, T., Shoji, I.
(2008). Virological characterization of the hepatitis C virus JFH-1 strain in lymphocytic cell lines. J. Gen. Virol.
89: 1587-1592
[Abstract]
[Full Text]
-
Song, Y., Friebe, P., Tzima, E., Junemann, C., Bartenschlager, R., Niepmann, M.
(2006). The Hepatitis C Virus RNA 3'-Untranslated Region Strongly Enhances Translation Directed by the Internal Ribosome Entry Site. J. Virol.
80: 11579-11588
[Abstract]
[Full Text]
-
Fok, V., Mitton-Fry, R. M., Grech, A., Steitz, J. A.
(2006). Multiple domains of EBER 1, an Epstein-Barr virus noncoding RNA, recruit human ribosomal protein L22. RNA
12: 872-882
[Abstract]
[Full Text]
-
Ivanyi-Nagy, R., Kanevsky, I., Gabus, C., Lavergne, J.-P., Ficheux, D., Penin, F., Fosse, P., Darlix, J.-L.
(2006). Analysis of hepatitis C virus RNA dimerization and core-RNA interactions.. Nucleic Acids Res
34: 2618-2633
[Abstract]
[Full Text]
-
Dutkiewicz, M., Ciesiolka, J.
(2005). Structural characterization of the highly conserved 98-base sequence at the 3' end of HCV RNA genome and the complementary sequence located at the 5' end of the replicative viral strand. Nucleic Acids Res
33: 693-703
[Abstract]
[Full Text]
-
Friebe, P., Boudet, J., Simorre, J.-P., Bartenschlager, R.
(2005). Kissing-Loop Interaction in the 3' End of the Hepatitis C Virus Genome Essential for RNA Replication. J. Virol.
79: 380-392
[Abstract]
[Full Text]
-
You, S., Stump, D. D., Branch, A. D., Rice, C. M.
(2004). A cis-Acting Replication Element in the Sequence Encoding the NS5B RNA-Dependent RNA Polymerase Is Required for Hepatitis C Virus RNA Replication. J. Virol.
78: 1352-1366
[Abstract]
[Full Text]
-
Lopez de Quinto, S., Saiz, M., de la Morena, D., Sobrino, F., Martinez-Salas, E.
(2002). IRES-driven translation is stimulated separately by the FMDV 3'-NCR and poly(A) sequences. Nucleic Acids Res
30: 4398-4405
[Abstract]
[Full Text]
-
Smith, R. M., Walton, C. M., Wu, C. H., Wu, G. Y.
(2002). Secondary Structure and Hybridization Accessibility of Hepatitis C Virus 3'-Terminal Sequences. J. Virol.
76: 9563-9574
[Abstract]
[Full Text]
-
Kim, M., Kim, H., Cho, S.-P., Min, M.-K.
(2002). Template Requirements for De Novo RNA Synthesis by Hepatitis C Virus Nonstructural Protein 5B Polymerase on the Viral X RNA. J. Virol.
76: 6944-6956
[Abstract]
[Full Text]
-
Friebe, P., Bartenschlager, R.
(2002). Genetic Analysis of Sequences in the 3' Nontranslated Region of Hepatitis C Virus That Are Important for RNA Replication. J. Virol.
76: 5326-5338
[Abstract]
[Full Text]
-
KIEFT, J.S., GRECH, A., ADAMS, P., DOUDNA, J.A.
(2001). Mechanisms of Internal Ribosome Entry in Translation Initiation. Cold Spring Harb Symp Quant Biol
66: 277-284
[Abstract]