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A Combinatorial Code for CPEB-Mediated c-myc Repression.

Koichi Ogami1,2, Keima Ogawa1, Shoko Sanpei1

  • 1Department of Biological Chemistry, Graduate School of Pharmaceutical Sciences, Nagoya City University, Nagoya 467-8603, Japan.

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|October 13, 2023
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Summary
This summary is machine-generated.

The cytoplasmic polyadenylation element-binding protein (CPEB) represses c-myc mRNA translation in somatic cells. A specific combination of consensus (cCPE) and non-consensus (ncCPE) elements dictates CPEB binding and c-myc mRNA decay.

Keywords:
CPEBRNA-binding proteinmRNA deadenylation

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Area of Science:

  • Molecular Biology
  • RNA Regulation
  • Gene Expression

Background:

  • Cytoplasmic polyadenylation element-binding protein (CPEB) regulates mRNA translation during embryonic development.
  • In non-neuronal somatic cells, CPEB's role shifts to repressing translation, including c-myc mRNA, via an unclear mechanism.
  • CPEB utilizes a combinatorial code of cytoplasmic polyadenylation elements (CPEs) for target mRNA regulation.

Purpose of the Study:

  • To elucidate the cis-regulatory mechanism of CPEB-mediated c-myc mRNA decay in somatic cells.
  • To define the combinatorial code of CPEs that governs CPEB binding and subsequent mRNA degradation.

Main Methods:

  • RNA mutagenesis was employed to identify critical CPE sequences.
  • Electrophoretic mobility shift assays (EMSAs) were used to quantify CPEB binding affinities.
  • mRNA degradation assays were performed to assess the functional impact of CPE combinations.

Main Results:

  • A combination of tandem consensus (cCPE) and non-consensus (ncCPE) elements forms a specific code for CPEB-mediated c-myc mRNA decay.
  • CPEB exhibits high-affinity binding to cCPEs (~250 nM) and low-affinity binding to ncCPEs (>900 nM).
  • Binding to cCPE enhances subsequent CPEB binding to ncCPE, with ncCPE being essential for degradation induction, further promoted by cCPE presence.

Conclusions:

  • A model is proposed where high-affinity CPEB binding to cCPE facilitates secondary binding to ncCPE.
  • This cooperative binding event leads to deadenylase recruitment, accelerated deadenylation, and repression of c-myc mRNA translation.
  • The combinatorial code of CPEs is crucial for regulating CPEB's function in mRNA decay.