CHARACTERIZATION OF THE DNA-BINDING DOMAIN OF THE AVIAN Y-BOX PROTEIN, CHKYB-2, AND MUTATIONAL ANALYSIS OF ITS SINGLE-STRAND BINDING MOTIF IN THE ROUS-SARCOMA VIRUS ENHANCER

chkYB-2 is a sequence-specific, single-stranded DNA binding chicken Y- box protein that promotes Rous sarcoma virus long terminal repeat (RSV LTR)-driven transcription in avian fibroblasts. The DNA-binding domai n of chkYB-2 has been mapped by characterizing the DNA binding propel ties of purified recombinant chkYB-2 mutant polypeptides. The data ind icate that the invariant cold shock domain (CSD) is necessary but not sufficient for association with DNA and suggest that another conserved region, adjacent to the carboxyl boundary of the CSD, plays a role in high-affinity DNA binding, chkYB-2 binds to a tandem repent of the 5' -GTACCACC-3' motif on the RSV LTR, Mutational analysis of this recogni tion sequence revealed the requirement of an essentially unaltered tem plate for both high-affinity binding by chkYB-2 as well as maximal tra nscriptional activity of the RSV LTR in vivo. The single-stranded DNA binding activity of chkYB-2 is augmented by Mg2+. The possible signifi cance of this finding for transactivation by a single-strand DNA bindi ng protein is discussed.