Cloning of a functional cDNA for the rabbit ferritin mRNA repressor protein. Demonstration of a tissue-specific pattern of expression

Abstract

Ferritin synthesis is controlled at the translational level in response to cellular iron status. A component of this regulatory system is the ferritin repressor protein (FRP) which binds to the iron-responsive element (IRE) located at the 5' end of all known ferritin mRNAs, thus inhibiting its translation. Antibodies against purified FRP were raised in mouse and used to isolate an FRP cDNA from a rabbit liver cDNA library cloned in the expression vector lambda gt11. The FRP cDNA encodes a 98.5-kilodalton protein which shares greater than 90% identity with IRE-binding proteins from other species. The FRP cDNA was placed under the transcriptional direction of the yeast GAL1 promoter. Yeast transformed with this gene express IRE-specific binding activity, illustrating the potential utility of yeast for the study of FRP structure/function. Analysis of FRP distribution in rabbit tissues shows that it is present in a variety of tissues. The levels of FRP differ dramatically from tissue to tissue, however. An examination of FRP mRNA levels and comparison to FRP protein suggest that synthesis of FRP is regulated transcriptionally and post-transcriptionally.