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mRNA with a <20-nt poly(A) tail imparted by the poly(A)-limiting element is translated as efficiently in vivo as long

Jing Peng1, Daniel R Schoenberg

  • 1Department of Molecular and Cellular Biochemistry, The Ohio State University, 1645 Neil Ave., Columbus, OH 43210-1218, USA.

RNA (New York, N.Y.)
|June 3, 2005
PubMed

Insights

The poly(A)-limiting element (PLE) restricts poly(A) tail length. It enhances translation of short poly(A) mRNA by substituting for poly(A)-binding protein (PABP) in cells.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • mRNA Metabolism

Background:

  • The poly(A) tail length is crucial for mRNA stability and translation efficiency.
  • The poly(A)-limiting element (PLE) is a conserved sequence element that limits poly(A) tail length to less than 20 nucleotides.
  • Poly(A)-binding protein (PABP) plays a key role in poly(A)-dependent translation.

Purpose of the Study:

  • To investigate the impact of the poly(A)-limiting element (PLE) on mRNA translation.
  • To compare the translation of PLE-containing mRNA with varying poly(A) tail lengths in different cellular contexts.
  • To elucidate the mechanism by which PLE influences translation, particularly in relation to PABP.

Main Methods:

  • Transfection of cells with PLE-containing and control mRNA.
  • In vitro translation assays using HeLa cell cytoplasmic extracts.
  • Manipulation of poly(A)-binding protein (PABP) levels and activity using poly(rA) and Paip2.
  • Analysis of mRNA accumulation, translation efficiency, and polysome binding.

Main Results:

  • In transfected cells, PLE-containing mRNA with short poly(A) tails accumulated 20% higher and translated as well as control mRNA with long poly(A) tails.
  • In vitro, PLE-containing mRNA showed reduced translation compared to controls with similar poly(A) tail lengths, indicating PLE did not overcome short poly(A) effects alone.
  • In vitro translation was enhanced when PABP activity was reduced (using poly(rA) or Paip2), and this effect was reversed by adding recombinant PABP, suggesting PLE acts by substituting for PABP.

Conclusions:

  • The poly(A)-limiting element (PLE) can enhance the translation of short poly(A) mRNA.
  • PLE's stimulatory effect on short poly(A) mRNA translation is dependent on the cellular context and PABP levels.
  • PLE appears to functionally substitute for PABP, thereby promoting the translation of mRNAs with limited poly(A) tail lengths.

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