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Structure and function analysis of the poliovirus cis-acting replication element (CRE).
Ian G Goodfellow1, David Kerrigan, David J Evans
1Faculty of Biomedical and Life Sciences, Division of Virology, University of Glasgow, Church Street, UK.
RNA (New York, N.Y.)
|January 30, 2003
Summary
The poliovirus cis-acting replication element (CRE) requires only a 14-nucleotide loop on a stable stem for VPg uridylylation. Its genomic location and duplication do not affect its function in picornavirus replication.
Area of Science:
- Virology
- Molecular Biology
- Structural Biology
Background:
- The poliovirus cis-acting replication element (CRE) is crucial for VPg protein uridylylation, a key step in viral genome replication.
- CRE is a conserved RNA element within the enterovirus polyprotein-coding region.
Purpose of the Study:
- To determine the native structure of the CRE.
- To identify critical regions of the CRE structure for its function.
- To investigate how genomic location influences CRE function.
Main Methods:
- Native structure determination of the CRE.
- Functional assays to define essential structural regions.
- Translocation and duplication experiments to assess genomic location effects.
Main Results:
- A 14-nucleotide unpaired terminal loop on a stable stem is sufficient for CRE activity.
- CRE function is maintained when translocated to the 5' noncoding region (3.7-kb away).
- CRE duplications do not impair viral replication.
Conclusions:
- The minimal functional unit of the CRE is a 14-nucleotide loop.
- CRE function is independent of its precise genomic location and can tolerate duplication.
- The precise mechanism of CRE coordination with 3CD(pro) remains to be elucidated, but direct interaction is not evident.