Publication Type:Journal Article
Source:RNA, Volume 6, Number 12, p.1808-20 (2000)
ISBN:1355-8382 (Print)<br/>1355-8382 (Linking)
Keywords:3' Untranslated Regions/ physiology, 5' Untranslated Regions/ physiology, Base Sequence, Cell-Free System, Consensus Sequence, Gene Expression Regulation, Viral, Luteovirus/ genetics, Models, Genetic, Molecular Sequence Data, Nucleic Acid Conformation, Peptide Chain Initiation, Translational/ physiology, Poly A/metabolism, Regulatory Sequences, Nucleic Acid, Ribosomes/ physiology, RNA Caps, RNA, Messenger/chemistry/ genetics, RNA, Viral/chemistry/ genetics, Structure-Activity Relationship, Triticum
Barley yellow dwarf virus RNA lacks both a 5' cap and a poly(A) tail, yet it is translated efficiently. It contains a cap-independent translation element (TE), located in the 3' UTR, that confers efficient translation initiation at the AUG closest to the 5' end of the mRNA. We propose that the TE must both recruit ribosomes and facilitate 3'-5' communication. To dissect its function, we determined the secondary structure of the TE and roles of domains within it. Nuclease probing and structure-directed mutagenesis revealed that the 105-nt TE (TE105) forms a cruciform secondary structure containing four helices connected by single-stranded regions. TE105 can function in either UTR in wheat germ translation extracts. A longer viral sequence (at most 869 nt) is required for full cap-independent translation in plant cells. However, substantial translation of uncapped mRNAs can be obtained in plant cells with TE105 combined with a poly(A) tail. All secondary structural elements and most primary sequences that were mutated are required for cap-independent translation in the 3' and 5' UTR contexts. A seven-base loop sequence was needed only in the 3' UTR context. Thus, this loop sequence may be involved only in communication between the UTRs and not directly in recruiting translational machinery. This structural and functional analysis provides a framework for understanding an emerging class of cap-independent translation elements distinguished by their location in the 3' UTR.
Guo, L<br/>Allen, E<br/>Miller, W A<br/>Research Support, Non-U.S. Gov't<br/>Research Support, U.S. Gov't, Non-P.H.S.<br/>United States<br/>RNA. 2000 Dec;6(12):1808-20.