title: Enhancement of protein production via the strong DIT1 terminator and two RNA-binding proteins in Saccharomyces cerevisiae.
abstract: Post-transcriptional upregulation is an effective way to increase the expression of transgenes and thus maximize the yields of target chemicals from metabolically engineered organisms. Refractory elements in the 3' untranslated region (UTR) that increase mRNA half-life might be available. In Saccharomyces cerevisiae, several terminator regions have shown activity in increasing the production of proteins by upstream coding genes; among these terminators the DIT1 terminator has the highest activity. Here, we found in Saccharomyces cerevisiae that two resident trans-acting RNA-binding proteins (Nab6p and Pap1p) enhance the activity of the DIT1 terminator through the cis element GUUCG/U within the 3'-UTR. These two RNA-binding proteins could upregulate a battery of cell-wall-related genes. Mutagenesis of the DIT1 terminator improved its activity by a maximum of 500% of that of the standard PGK1 terminator. Further understanding and improvement of this system will facilitate inexpensive and stable production of complicated organism-derived drugs worldwide.
authors.affiliation: Toyota Central R&D Labs. Inc., Nagakute, Aichi 480-1192, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Systems Biochemistry in Pathology and Regeneration, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Kitagawa
authors.affiliation: Toyota Central R&D Labs. Inc., Nagakute, Aichi 480-1192, Japan.
authors.forename: Mamoru
authors.initials: M
authors.lastname: Yamanishi
authors.affiliation: Toyota Central R&D Labs. Inc., Nagakute, Aichi 480-1192, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Katahira
authors.affiliation: Laboratory of Microbial Technology, Graduate School of Science and Technology, Kyoto Institute of Technology, Kyoto 606-8585, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Izawa
authors.affiliation: Laboratory of Molecular Cell Biology, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Irie
authors.affiliation: Systems Biochemistry in Pathology and Regeneration, Yamaguchi University Graduate School of Medicine, Ube, Yamaguchi 755-8505, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Furutani-Seiki
authors.affiliation: Toyota Central R&D Labs. Inc., Nagakute, Aichi 480-1192, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsuyama
mesh_terms: D020413:3' Untranslated Regions; D015870:Gene Expression; D019877:Hydroxymethyl and Formyl Transferases / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D013728:Terminator Regions, Genetic
chemical_list: D020413:3' Untranslated Regions; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C503580:DIT1 protein, S cerevisiae; D019877:Hydroxymethyl and Formyl Transferases