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Cloning and characterization of seven human forkhead proteins: binding site specificity and DNA bending.
Author(s) -
Pierrou S.,
Hellqvist M.,
Samuelsson L.,
Enerbäck S.,
Carlsson P.
Publication year - 1994
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1994.tb06827.x
Subject(s) - biology , forkhead transcription factors , dna binding site , gene , genetics , dna , oligonucleotide , complementary dna , dna binding protein , binding site , hmg box , dna binding domain , microbiology and biotechnology , transcription factor , gene expression , promoter
The forkhead domain is a monomeric DNA binding motif that defines a rapidly growing family of eukaryotic transcriptional regulators. Genetic and biochemical data suggest a central role in embryonic development for genes encoding forkhead proteins. We have used PCR and low stringency hybridization to isolate clones from human cDNA and genomic libraries that represent seven novel forkhead genes, freac‐1 to freac‐7. The spatial patterns of expression for the seven freac genes range from specific for a single tissue to nearly ubiquitous. The DNA binding specificities of four of the FREAC proteins were determined by selection of binding sites from random sequence oligonucleotides. The binding sites for all four FREAC proteins share a core sequence, RTAAAYA, but differ in the positions flanking the core. Domain swaps between two FREAC proteins identified two subregions within the forkhead domain as responsible for creating differences in DNA binding specificity. Applying a circular permutation assay, we show that binding of FREAC proteins to their cognate sites results in bending of the DNA at an angle of 80‐90 degrees.