title: Efficient transgene expression by alleviation of translational repression in plant cells.
abstract: Global translational repression under abiotic stress influences translation of both endogenous and transgene mRNAs. Even in plant cell culture, hypoxia and nutrient deficient stress arise during the growth process. In this study, we first demonstrated the existence of global translational repression in Arabidopsis T87 cultured cells over a time course following inoculation. Next, we performed genome-wide analysis, which revealed that the translational states of endogenous mRNAs differed significantly between growth and stationary phase cells. This analysis showed that translation from most mRNAs was repressed upon stationary phase. Otherwise, a part of mRNA including alcohol dehydrogenase (ADH) gene was recalcitrant to the repression. Furthermore, by polysome analysis and followed quantitative reverse transcription PCR analysis of transformants having 5'untranslated regions (UTRs) of ADH or translationally repressed At3g47610 mRNA fused to reporter gene, we demonstrated that polysomal associations of reporter mRNAs were in accordance with those the mRNAs from which their 5'UTR derived, suggesting that the 5'UTR is an important determinant of the translational state of mRNAs in stationary phase cells. Finally, we demonstrated the effectiveness of 5'UTR of ADH mRNA in transformants derived from the BY-2 tobacco cell line. These results suggested that 5'UTR of ADH mRNA would be a useful element for efficient transgene expression upon stationary phase.
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Ueda
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Renya
authors.initials: R
authors.lastname: Okawara
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Shotaro
authors.initials: S
authors.lastname: Yamasaki
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Sanada
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Eri
authors.initials: E
authors.lastname: Kinoshita
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Arata
authors.initials: A
authors.lastname: Yoneda
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Taku
authors.initials: T
authors.lastname: Demura
authors.affiliation: Graduate School of Biological Science, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan. Electronic address: kou@bs.naist.jp.
authors.forename: Ko
authors.initials: K
authors.lastname: Kato
mesh_terms: D020121:5' Untranslated Regions; D000426:Alcohol Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D017360:Arabidopsis / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D059828:Plant Cells / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011132:Polyribosomes / Q000235:genetics; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D019076:Transgenes
chemical_list: D020121:5' Untranslated Regions; D010940:Plant Proteins; D012333:RNA, Messenger; D000426:Alcohol Dehydrogenase
keywords: 5′UTR; Abiotic stress; Plant cell cultures; Recombinant proteins; Transgenic; Translation; Translational repression