THE HMG BOX OF SRY IS A CALMODULIN-BINDING DOMAIN

The HMG box domain of the testis determining factor, SRY, includes a b asic amphiphilic sequence common to calmodulin (CaM) binding proteins, By affinity chromatography, native gel electrophoresis and fluorescen ce spectroscopy, we show the calcium-dependent binding of SRY to CaM, Binding occurs via the HMG box and an SRY peptide of residues 57-80 bi nds CaM like the intact domain, SRY/CaM complex formation is specifica lly inhibited by the SRY DNA binding site sequence, AACAAT, but not a mutated sequence, Fluorescence spectra of the SRY/CaM complex indicate 1:1 stoichiometry and that binding is accompanied by a conformational change in SRY, The A domain of HMG1 also binds CaM and me propose tha t CaM binding is a property of the wider HMG box family, including SOX and TCF/LEF proteins, These results suggest that CaM may regulate the DNA binding activity of HMG box transcription factors.