Purification and characterization of winter flounder antifreeze peptide messenger ribonucleic acid

The serum of winter flounder contains a group of small antifreeze peptides which lower the freezing point of their body fluids during the winter months. The poly(A)-containing mRNA coding for these peptides has been isolated from livers of the winter specimens. When the isolated antifreeze mRNA was analyzed by a denaturing polyacrylamide gel electrophoresis, at least two distinct bands approximately 450 nucleotides in length are visible. In a wheat germ cell-free protein synthetic system these mRNAs direct the synthesis of small peptides which can be precipitated by antisera against purified winter flounder antifreeze peptides. Full-length cDNA was synthesized from the isolated antifreeze mRNA by avian myeloblastosis reverse transcriptase. From the RNA excess hybridization kinetic analysis, there are probably three different mRNAs coding for the antifreeze peptides. Using the radioactive cDNA probe, it was estimated that 1% of the total RNA in liver of a January specimen is antifreeze mRNA. RNA from a summar specimen showed no significant hybridization even at high concentrations of RNA. These results indicate that the control of antifreeze peptide biosynthesis relies at least in part on the synthesis or degradation of translatable mRNA.