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Two cellular proteins bind specifically to a purine-rich sequence necessary for the destabilization function of a

C Y Chen1, Y You, A B Shyu

  • 1Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77030.

Molecular and Cellular Biology
|December 1, 1992
PubMed

Insights

Two proteins bind a purine-rich region in c-fos mRNA, targeting it for rapid degradation. This interaction is crucial for the mRNA instability element

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • RNA Metabolism

Background:

  • The c-fos proto-oncogene mRNA is rapidly degraded in mammalian fibroblasts after growth factor stimulation.
  • This rapid decay is mediated by specific sequence elements within the mRNA.
  • A key instability determinant resides in a 0.32-kb sequence within the protein-coding region.

Purpose of the Study:

  • To identify and characterize protein factors that interact with the c-fos mRNA instability element.
  • To elucidate the mechanism by which the c-fos coding region determinant of mRNA instability (CRDI) functions.
  • To investigate the evolutionary conservation of the functional elements within the c-fos mRNA.

Main Methods:

  • Gel mobility shift assays to detect protein-DNA interactions.
  • UV cross-linking experiments to identify bound proteins.
  • Sequence analysis and comparison across the fos gene family.
  • Functional assays involving deletion of specific RNA regions.

Main Results:

  • Two distinct protein factors specifically bind to a 56-nucleotide purine-rich sequence at the 5' end of the 0.32-kb c-fos CRDI.
  • One protein factor is polysome-associated, while the other is in the post-ribosomal supernatant.
  • The purine-rich sequence is conserved in fos gene family members.
  • Deletion of this purine-rich region significantly impairs the RNA-destabilizing function of the CRDI.

Conclusions:

  • The binding of two specific proteins to the purine-rich sequence is implicated in the rapid decay of c-fos mRNA.
  • This protein-RNA interaction is a key mechanism underlying the function of the c-fos CRDI.
  • The conserved nature of this sequence suggests its fundamental importance in regulating c-fos expression.

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