Publication Type:Journal Article
Source:BMC Biotechnol, Volume 12, p.22 (2012)
ISBN:1472-6750 (Electronic)<br/>1472-6750 (Linking)
Keywords:3' Untranslated Regions/ genetics, 5' Untranslated Regions/ genetics, Avena sativa/metabolism, Genes, Reporter, Genome, Viral, Luteovirus/genetics, Plant Cells/metabolism, Protein Biosynthesis/genetics, Protoplasts/metabolism, RNA, Messenger/metabolism, RNA, Viral/ genetics/metabolism, Tobacco necrosis satellite virus/genetics, Tobacco/metabolism, Tobamovirus/genetics, Tombusvirus/genetics, Triticum/metabolism
BACKGROUND: Whole plants or plant cell cultures can serve as low cost bioreactors to produce massive amounts of a specific protein for pharmacological or industrial use. To maximize protein expression, translation of mRNA must be optimized. Many plant viral RNAs harbor extremely efficient translation enhancers. However, few of these different translation elements have been compared side-by-side. Thus, it is unclear which are the most efficient translation enhancers. Here, we compare the effects of untranslated regions (UTRs) containing translation elements from six plant viruses on translation in wheat germ extract and in monocotyledenous and dicotyledenous plant cells. RESULTS: The highest expressing uncapped mRNAs contained viral UTRs harboring Barley yellow dwarf virus (BYDV)-like cap-independent translation elements (BTEs). The BYDV BTE conferred the most efficient translation of a luciferase reporter in wheat germ extract and oat protoplasts, while uncapped mRNA containing the BTE from Tobacco necrosis virus-D translated most efficiently in tobacco cells. Capped mRNA containing the Tobacco mosaic virus omega sequence was the most efficient mRNA in tobacco cells. UTRs from Satellite tobacco necrosis virus, Tomato bushy stunt virus, and Crucifer-infecting tobamovirus (crTMV) did not stimulate translation efficiently. mRNA with the crTMV 5' UTR was unstable in tobacco protoplasts. CONCLUSIONS: BTEs confer the highest levels of translation of uncapped mRNAs in vitro and in vivo, while the capped omega sequence is most efficient in tobacco cells. These results provide a basis for understanding mechanisms of translation enhancement, and for maximizing protein synthesis in cell-free systems, transgenic plants, or in viral expression vectors.
Fan, Qiuling<br/>Treder, Krzysztof<br/>Miller, W Allen<br/>2R01-GM067104/GM/NIGMS NIH HHS/United States<br/>R01 GM067104/GM/NIGMS NIH HHS/United States<br/>Research Support, N.I.H., Extramural<br/>Research Support, Non-U.S. Gov't<br/>England<br/>BMC Biotechnol. 2012 May 6;12:22. doi: 10.1186/1472-6750-12-22.