Structure of the rabbit cytochrome P450IIC3 gene, a constitutive member of the P450IIC subfamily.
Fragments of rabbit DNA have been cloned which encompass the gene for rabbit cytochrome P450IIC3. Cytochrome P450IIC3 is a constitutive member of the cytochrome P450IIC subfamily which contains both constitutive and phenobarbital-responsive genes. The cytochrome P450IIC3 gene spans at least 25 kbp and contains 8 introns which have 5'GT's and 3'AG's and are located in the same positions as in other family II genes. The 5' flanking region contains a consensus TATA site about 25 bp from the RNA initiation site which was mapped by the primer extension method. Other potential regulatory sequences include a CCAAT sequence and sequences similar to binding sites for the liver-specific factor HNF-1 and the general transcription factors AP-1 and OCT. The gene sequence differs from the cytochrome P450IIC3 cloned cDNA sequence in only 2 of 1400 nucleotides, 1 of which results in an amino acid change. Since sequences coding for exon 1 and part of exon 2 were not present in the cloned cytochrome P450IIC3 cDNA, a 475-nucleotide fragment of the 5' end of the cDNA was amplified by the polymerase chain reaction, cloned, and sequenced. The sequence of this cDNA was identical with the gene exon sequence. The protein sequence derived from the gene differs in nine positions from that determined directly for the protein. A comparison of the amino acid sequence by exons with other cytochrome P450IIC proteins revealed that, in general, the similarity was greater in C-terminal exons than in N-terminal ones.(ABSTRACT TRUNCATED AT 250 WORDS)
Chan
G
G
Department of Physiology and Biophysics, University of Illinois, Urbana-Champaign 61801.
Kemper
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D004247:DNA / Q000032:analysis; D005091:Exons; D007438:Introns; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits
D012333:RNA, Messenger; D004247:DNA; D003577:Cytochrome P-450 Enzyme System