SCHISTOSOMA-MANSONI - INTERACTION OF NUCLEAR EXTRACTS WITH THE CCAAT-BINDING SITE REVEALED BY THE GEL SHIFT ASSAY

Analysis of the 5'-flanking region of the gene encoding the 28-kDa g]u tathione S-transferase of Schistosoma mansoni (Sm28GST indicated the p resence of motifs identical to AP-1 and CCAAT-family transcription fac tor recognition sequences. Gel retardation experiments showed that nuc lear extracts from adult S. mansoni bound to an oligodeoxynucleotide c ontaining at CCAAT box. A DNA fragment corresponding to the region of Sm28GST containing the CCAAT motif was demonstrated to interact with s chistosome nuclear proteins. This binding was dependent on the presenc e of the CCAAT pentanucleotide motif. Nuclear factor Y (NF-Y) is a mem ber of the CCAAT transcription factor family that has absolute require ment for the CCAAT sequence and that is highly conserved throughout ev olution. The results of a PCR-based strategy aimed at cloning the NF-Y A protein of S. mansoni are presented. (C) 1995 Academic Press, Inc.