title: Enhancing recombinant protein production in human cell lines with a constitutive transport element and mRNA export proteins.
abstract: Recent research into mRNA maturation processes in the nucleus has identified a number of proteins involved in mRNA transcription, capping, splicing, end processing and export. Among them, the Tap-p15 heterodimer acts as an mRNA export receptor. Tap-p15 is recruited onto fully processed mRNA in the nucleus, which is ready for export to the cytoplasm, through associating with Aly or SR proteins on mRNA, or by directly associating with a constitutive transport element (CTE), an RNA element derived from type D retroviruses. mRNA containing a CTE is exported to the cytoplasm by directly associating with Tap-p15, even in the absence of Tap-recruiting proteins such as Aly or SR proteins on the mRNA. Here, we showed that the use of a CTE enhanced the expression of recombinant protein in human cell lines. The co-expression of reporter proteins and Tap-p15 also enhanced recombinant protein expression. Moreover, the use of a CTE and Tap-p15 synergistically further enhanced the recombinant protein expression. In addition to Tap-p15, several Tap-p15-recruiting proteins, including Aly and SR proteins, enhanced recombinant protein expression, albeit independently of the CTE. The incorporation of a CTE and Tap-p15-recruiting proteins into protein expression system is useful to increase recombinant protein yield in human cells.
authors.affiliation: Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto 606-8502, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Aihara
authors.forename: Naoko
authors.initials: N
authors.lastname: Fujiwara
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Yamazaki
authors.forename: Taiho
authors.initials: T
authors.lastname: Kambe
authors.forename: Masaya
authors.initials: M
authors.lastname: Nagao
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Hirose
authors.forename: Seiji
authors.initials: S
authors.lastname: Masuda
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015153:Blotting, Western; D045744:Cell Line, Tumor; D004921:Erythropoietin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D029543:Nucleocytoplasmic Transport Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D034443:RNA Transport; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D038621:Regulatory Sequences, Ribonucleic Acid
chemical_list: C498546:NXT1 protein, human; D029543:Nucleocytoplasmic Transport Proteins; D011993:Recombinant Fusion Proteins; D038621:Regulatory Sequences, Ribonucleic Acid; D004921:Erythropoietin; D008156:Luciferases; D000469:Alkaline Phosphatase