Characterization of the chicken fatty acid synthase gene 5' part and promoter region.
Fatty acid synthase activity has been shown to be regulated mainly at the transcriptional level under both dietary and hormonal influences. As a first step towards elucidating the factors involved, we isolated and characterized chicken genomic clones encompassing the 5' part of the chicken fatty acid synthase gene and its flanking region. The entire region of the cloned DNA spans 30 kb, and the first three exons of the gene were mapped to a 6.3-kb genomic fragment. The transcription initiation site was determined after subcloning the cDNA which encodes the 5' end of the mRNA. The first exon, which was 129 bp long, was located approximately 5.3 kb upstream of the second exon, which contained the start codon. In the 5' flanking region, putative TATA and CAAT boxes were located 30 and 92 bp, respectively, upstream of the transcription initiation site. The 5' flanking region contained numerous sequences corresponding to consensus binding sites for transcription factors. Various lengths of flanking sequences extending up to 1028 bp upstream of the transcription initiation site and containing 100 bp of the first exon were linked to the bacterial chloramphenicol acetyltransferase gene; in this study, these constructs were analyzed in transient transfection assays in human hepatoma cells. The proximal 125-bp sequence upstream of the transcription start site was shown to be a basal promoter. The cloning and characterization of the chicken fatty-acid synthase gene provides some further insight into the regulation of fatty acid synthesis in birds as compared to mammals.
Le Fur
N
N
Laboratoire de Génétique Animale, INRA/ENSAR, Rennes, France.
el Khadir-Mounier
C
C
Powell
R S
RS
Diot
C
C
Mallard
J
J
Douaire
M
M
D000818:Animals; D001483:Base Sequence; D002645:Chickens / Q000235:genetics*; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism; D003001:Cloning, Molecular; D018387:Codon, Initiator / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D064429:Fatty Acid Synthases / Q000235:genetics*; D017930:Genes, Reporter; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D015183:Restriction Mapping; D012260:Ribonucleases / Q000378:metabolism; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D018387:Codon, Initiator; D018076:DNA, Complementary; D012333:RNA, Messenger; D015500:Chloramphenicol O-Acetyltransferase; D064429:Fatty Acid Synthases; D012260:Ribonucleases