Genomic structure and promoter analysis of the mouse neutral ceramidase gene.
We report here the molecular cloning of the mouse neutral ceramidase gene and its promoter analysis. The gene, composed of 27 exons ranging in size from 40 to 292 bp, spans more than 70 kb. Analysis of the 5(')-flanking region of the ceramidase genes revealed that the first exon of the gene of mouse liver was exactly the same as that of mouse kidney and Swiss 3T3 fibroblasts but completely different from that of mouse brain. The putative promoter regions of liver and brain ceramidase genes contained several well-characterized promoter elements such as GATA-2, C/EBP, and HNF3beta but lacked TATA and CAAT boxes, a typical feature of a housekeeping gene, although the expression is regulated in a tissue-specific manner. Interestingly, a GC box was exclusively found in the putative promoter of mouse liver whereas potential AP1 and AP4 binding sites were present in that of mouse brain. By a luciferase reporter gene assay, it was shown that the GC-rich region, which exists just upstream of the first exon, conferred the promoter activity in Swiss 3T3 cells.
Okino
Nozomu
N
Department of Bioscience and Biotechnology, Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, 812-8581, Fukuoka, Japan.
Mori
Kaoru
K
Ito
Makoto
M
D016475:3T3 Cells; D000581:Amidohydrolases / Q000235:genetics* / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology; D055572:Ceramidases; D004247:DNA / Q000378:metabolism; D008099:Liver / Q000201:enzymology; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D008969:Molecular Sequence Data; D055576:Neutral Ceramidase; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011485:Protein Binding
D004247:DNA; D008156:Luciferases; D000581:Amidohydrolases; C526747:Asah2 protein, mouse; D055572:Ceramidases; D055576:Neutral Ceramidase