Translation of overlapping open reading frames promoted by type 2 IRESs in avian calicivirus genomes

Description

Primary data (gel scans) Figure 3. The mechanism of initiation on the grey teal calicivirus (GTCV) IRES. Figure 4. Initiation factor requirements for initiation on the RaCV1 IRES. Figure 5. Influence of AUG codons and structural elements inserted downstream of the Yn-Xm-AUG motif on RaCV1 IRES-mediated initiation of translation. Figure 6. Requirement for subunits of eIF4F for initiation on the RaCV1 IRES. Figure 7. Influence of the GNRA tetraloop at the apex of subdomain Ib on RaCV1 IRES function. Figure 8. Interaction of Ebp1 with type 2 IRESs. Figure 9. Influence of mutations in RaCV1 IRES subdomain Id on binding of Ebp1 assessed by directed hydroxyl radical probing and IRES activity.

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Translation of calicivirus mRNAs was assayed using rabbit reticulocyte lysate. The mechanism of initiation was determined by in vitro reconstitution, using individual purified native and/or recombinant components of the translation apparatus (ribosomal subunits, initiation and elongation factors, IRES trans-acting factors, aminoacyl-tRNAs, mRNAs). Interactions of factors with calicivirus mRNAs were mapped using Fe(II)BABE-modified proteins in directed hydroxyl radical probing experiments.

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