DOI: 10.1128/jvi.76.10.5024-5033.2002. Pestivirus Internal Ribosome Entry Site (IRES) Structure and Function: Elements in the 5′ Untranslated Region Important for IRES Function

Pestivirus Internal Ribosome Entry Site (IRES) Structure and Function: Elements in the 5′ Untranslated Region Important for IRES Function

10.1128/jvi.76.10.5024-5033.2002

Crossref journal-article

American Society for Microbiology

Journal of Virology (235)

Abstract

ABSTRACT The importance of certain structural features of the 5′ untranslated region of classical swine fever virus (CSFV) RNA for the function of the internal ribosome entry site (IRES) was investigated by mutagenesis followed by in vitro transcription and translation. Deletions made from the 5′ end of the CSFV genome sequence showed that the IRES boundary was close to nucleotide 65: thus, the IRES includes the whole of domain II but no sequences upstream of this domain. Deletions which invaded domain II even to a small extent reduced activity to about 20% that of the full-length structure, and this 20% residual activity persisted with more extensive deletions until the whole of domain II had been removed and the deletions invaded the pseudoknot, whereupon IRES activity fell to zero. The importance of both stems of the pseudoknot was verified by making mutations in both sides of each stem; this severely reduced IRES activity, but the compensating mutations which restored base pairing caused almost full IRES function to be regained. The importance of the length of the loop linking the two stems of the pseudoknot was demonstrated by the finding that a reduction in length from the wild-type AUAAAAUU to AUU almost completely abrogated IRES activity. Random A→U substitutions in the wild-type sequence showed that IRES activity was fairly proportional to the number of A residues retained in this pseudoknot loop, with a preference for clustered neighboring A residues rather than dispersed As. Finally, it was found that the sequence of the highly conserved domain IIIa loop is, rather surprisingly, not important for the maintenance of full IRES activity, although amputation of the entire domain IIIa stem and loop was highly debilitating. These results are interpreted in the light of recent models, derived from cryo-electron microscopy, of the interaction of the closely related hepatitis C virus IRES with 40S ribosomal subunits.

Bibliography

Fletcher, S. P., & Jackson, R. J. (2002). Pestivirus Internal Ribosome Entry Site (IRES) Structure and Function: Elements in the 5â² Untranslated Region Important for IRES Function. Journal of Virology, 76(10), 5024â5033.

Authors 2

Simon P. Fletcher (first)
Richard J. Jackson (additional)

References 36 Referenced 96

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Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 6:09 a.m.)
Deposited	3 years, 6 months ago (March 5, 2022, 1:53 a.m.)
Indexed	1 month, 2 weeks ago (July 20, 2025, 12:21 a.m.)
Issued	23 years, 3 months ago (May 15, 2002)
Published	23 years, 3 months ago (May 15, 2002)
Published Print	23 years, 3 months ago (May 15, 2002)

Funders 0

None

BibTeX

@article{Fletcher_2002, title={Pestivirus Internal Ribosome Entry Site (IRES) Structure and Function: Elements in the 5′ Untranslated Region Important for IRES Function}, volume={76}, ISSN={1098-5514}, url={http://dx.doi.org/10.1128/jvi.76.10.5024-5033.2002}, DOI={10.1128/jvi.76.10.5024-5033.2002}, number={10}, journal={Journal of Virology}, publisher={American Society for Microbiology}, author={Fletcher, Simon P. and Jackson, Richard J.}, year={2002}, month=may, pages={5024–5033} }

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