This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Graff, J.
Right arrow Articles by Ehrenfeld, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Graff, J.
Right arrow Articles by Ehrenfeld, E.

 Previous Article  |  Next Article 

J Virol, May 1998, p. 3571-3577, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Coding Sequences Enhance Internal Initiation of Translation by Hepatitis A Virus RNA In Vitro

Judith Graff* and Ellie Ehrenfelddagger

Department of Molecular Biology and Biochemistry, University of California---Irvine, Irvine, California 92697

Received 6 October 1997/Accepted 3 February 1998

Hepatitis A virus (HAV), unlike other picornaviruses, has a slow-growth phenotype in permissive cell lines and in general does not induce host cell cytopathology. Although there are no published reports of productive infection of HeLa cells by HAV, HAV RNA appears to be readily translated in HeLa cells when transcribed by T7 RNA polymerase provided by a recombinant vaccinia virus. The 5' noncoding region of HAV was fused to poliovirus (PV) coding sequences to determine the effect on translation efficiency in HeLa cell extracts in vitro. Conditions were optimized for utilization of the HAV internal ribosome entry segment (IRES). Transcripts from chimeric constructs fused precisely at the initiation codon were translated very poorly. However, chimeric RNAs which included 114 or more nucleotides from the HAV capsid coding sequences downstream of the initiation codon were translated much more efficiently than those lacking these sequences, making HAV-directed translation efficiency similar to that directed by the PV IRES. Sixty-six nucleotides were insufficient to confer increased translation efficiency. The most 5'-terminal HAV 138 nucleotides, previously determined to be upstream of the IRES, had an inhibitory effect on translation efficiency. Constructs lacking these terminal sequences, or those in which the PV 5'-terminal sequences replaced those from HAV, translated three- to fourfold better than those with the intact HAV 5'-terminal end.

* Corresponding author. Present address: National Institutes of Health, NIAID, LID, Hepatitis Virus Section, Building 7, Room 200, 7 Center Dr., MSC 0740, Bethesda, MD 20892-0740. Phone: (301) 496-6227. Fax: (301) 402-0524. E-mail: jgraff{at}atlas.niaid.nih.gov.

dagger Present address: Center for Scientific Review, National Institutes of Health, Bethesda, MD 20892.

J Virol, May 1998, p. 3571-3577, Vol. 72, No. 5
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Dobrikova, E. Y., Grisham, R. N., Kaiser, C., Lin, J., Gromeier, M. (2006). Competitive Translation Efficiency at the Picornavirus Type 1 Internal Ribosome Entry Site Facilitated by Viral cis and trans Factors.. J. Virol. 80: 3310-3321 [Abstract] [Full Text]  
  • Piron, M., Beguiristain, N., Nadal, A., Martinez-Salas, E., Gomez, J. (2005). Characterizing the function and structural organization of the 5' tRNA-like motif within the hepatitis C virus quasispecies. Nucleic Acids Res 33: 1487-1502 [Abstract] [Full Text]  
  • Yi, M., Lemon, S. M. (2002). Replication of Subgenomic Hepatitis A Virus RNAs Expressing Firefly Luciferase Is Enhanced by Mutations Associated with Adaptation of Virus to Growth in Cultured Cells. J. Virol. 76: 1171-1180 [Abstract] [Full Text]  
  • Tang, S., Collier, A. J., Elliott, R. M. (1999). Alterations to both the Primary and Predicted Secondary Structure of Stem-Loop IIIc of the Hepatitis C Virus 1b 5' Untranslated Region (5'UTR) Lead to Mutants Severely Defective in Translation Which Cannot Be Complemented in trans by the Wild-Type 5'UTR Sequence. J. Virol. 73: 2359-2364 [Abstract] [Full Text]  
  • Sasaki, J., Nakashima, N. (1999). Translation Initiation at the CUU Codon Is Mediated by the Internal Ribosome Entry Site of an Insect Picorna-Like Virus In Vitro. J. Virol. 73: 1219-1226 [Abstract] [Full Text]  
  • Graff, J., Cha, J., Blyn, L. B., Ehrenfeld, E. (1998). Interaction of Poly(rC) Binding Protein 2 with the 5' Noncoding Region of Hepatitis A Virus RNA and Its Effects on Translation. J. Virol. 72: 9668-9675 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»