Identification of proteins involved in the regulation of yeast iso- 1-cytochrome C expression by oxygen.

On the basis of a gel electrophoresis retardation assay, protein(s) which interact specifically with the upstream activating site (UASc) of the yeast iso-1-cytochrome C (CYC1) gene were identified and separated by heparin ultrogel chromatography. DNase I protection experiments indicate that these factors protect a 23-bp sequence overlapping the UASc site previously defined. The specific binding activity is strongly reduced in extracts prepared from a wild-type strain grown anaerobically. It is absent in a mutant strain blocked in the biosynthesis of heme but it is restored upon the addition of the missing precursor, delta amino levulinic acid (dALA) to the growth medium. In contrast, the binding activity does not differ significantly in extracts form a wild-type strain grown in either glucose or glycerol as carbon source. These data strongly argue that the CYC1 UAS binding protein(s) that we have identified mediate the oxygen and heme control of cytochrome C biosynthesis.