Reconstitution of yeast translation elongation and termination in vitro utilizing CrPV IRES-containing mRNA

• Taisho Abe

• Riku Nagai

• Hiroaki Imataka

• Nono Takeuchi-Tomita

Abstract

We developed an in vitro translation system from yeast, reconstituted with purified translation elongation and termination factors and programmed by CrPV IGR IRES-containing mRNA, which functions in the absence of initiation factors. The system is capable of synthesizing the active reporter protein, nanoLuciferase, with a molecular weight of 19 kDa. The protein synthesis by the system is appropriately regulated by controlling its composition, including translation factors, amino acids and antibiotics. We found that a high eEF1A concentration relative to the ribosome concentration is critically required for efficient IRES-mediated translation initiation, to ensure its dominance over IRES-independent random internal translation initiation.

Journal

• The journal of biochemistry

  The journal of biochemistry 167 (5), 441-450, 2020-05

  Tokyo : Japanese Biochemical Society

Keywords

• CrPV IGR IRES
• in vitro translation
• translation elongation
• translation termination
• yeast

Details

• CRID: 1522825130416649728
• NII Article ID: 40022226784
• NII Book ID: AA00694073
• ISSN: 0021924X
• ISSN: 17562651
• DOI: 10.1093/jb/mvaa021
• NDL BIB ID: 030402958
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I030402958
• Web Site: http://academic.oup.com/jb/advance-article-pdf/doi/10.1093/jb/mvaa021/32916015/mvaa021.pdf
• Web Site: http://academic.oup.com/jb/article-pdf/167/5/441/33466228/mvaa021.pdf
• Web Site: https://search.jamas.or.jp/link/ui/2021087558
• Text Lang: en
• NDL Source Classification:
  • ZR2(科学技術--生物学--生化学)
• Data Source:
  • NDL
  • Crossref
  • CiNii Articles
  • KAKEN