Translation initiation from the ribosomal A site or the P site, dependent on the conformation of RNA pseudoknot I in dicistrovirus RNAs
Author(s)
Kamoshita, Nobuhiko; Nomoto, Akio; Rajbhandary, Uttam L
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Translation initiation of the second ORF of insect dicistrovirus RNA depends on an internal ribosomal entry site (IRES) in its intergenic region (IGR) and is exceptional in using a codon other than AUG and in not using the canonical initiator methionine tRNA. Studies in vitro suggest that pseudoknot I (PKI) immediately preceding the initiation codon occupies the ribosomal P site and that an elongator tRNA initiates translation from the ribosomal A site. Using dicistronic reporters carrying mutations in the initiation codon of the second ORF and mutant elongator or initiator tRNAs capable of reading these codons, we provide direct evidence for initiation from the A site in mammalian cells and, under certain conditions, also from the P site. Initiation from the A but not the P site requires PKI. Thus, PKI structure may be dynamic, and optimal IGR IRES-mediated translation of dicistroviral RNAs may require trans-acting factors to stabilize PKI.
Description
available in PMC 2010 July 31.
Date issued
2009-07Department
Massachusetts Institute of Technology. Department of BiologyJournal
Molecular Cell
Publisher
Elsevier B.V.
Citation
Kamoshita, Nobuhiko, Akio Nomoto, and Uttam L. RajBhandary. “Translation Initiation from the Ribosomal A Site or the P Site, Dependent on the Conformation of RNA Pseudoknot I in Dicistrovirus RNAs.” Molecular Cell 35.2 (2009): 181–190. Web.
Version: Author's final manuscript