ISOLATION AND FUNCTIONAL-ANALYSIS OF THE PROMOTER OF THE RAT CMP-NEUAC-GM3 ALPHA-2,8 SIALYLTFANSFERASE GENE

A 2.1-kb 5'-flanking fragment of the rat CMP-NeuAc:GM3 alpha 2,8 sialy ltransferase (GD3-synthase) gene was cloned by the genomic walking pro cedure. The promoter activity of the fragment was assessed in F-II cel ls by transient transfection and the locations for the basal and maxim al promoter activities were defined. Primer extension analysis identif ied a transcription start site approximately 98 bp upstream of the ATG start codon. DNA sequence analysis of the promoter revealed a number of consensus binding sites for known transcription factors such as SP1 . AP1. NF kappa B, C/EBP and TFIID, and a repeat GC-GT sequence motif seen for the formation of Z-type DNA. Both TATA and CCAAT boxes were n ot found in the promoter. Our results from deletion constructs suggest ed that both positive and negative cis-acting regulatory regions were present in this TATA-less promoter of the rat GD3-synthase gene. (C) 1 998 Elsevier Science B.V.