Retinoic acid metabolizing cytochrome P450
The present invention provides a novel retinoic acid metabolizing cytochrome P450, P450RAI-3, that is predominantly expressed in the adrenal gland. Methods for and uses of the new polynucleotide, polypeptide, fragments thereof and modulators thereof, include the treatment of cancer.
1. An isolated cDNA molecule comprising a sequence selected from the group consisting of: (a) SEQ. ID. NO. 10 or encoding the amino acid sequence of SEQ. ID. NO. 11; (b) a polynucleotide of (a) wherein T can also be U: (c) a polynucleotide having a nucleic acid sequence which differs from any of the nucleic acid molecules of (a) to (c) in codon due to the degeneracy of the genetic code; (d) a polynucleotide that is a variant, of any one of the polynucleotides of (a) to (c); wherein said variant is at least 95% identical to SEQ ID NO:10 and encodes a polypeptide that retains the substrate specificity of the polypeptide comprising the amino acid sequence of SEQ ID NO:11 and (e) a polynucleotide that is a fragment of any one of the polynucleotides of (a) to (d), wherein said fragment encodes a polypeptide that retains the substrate specificity of the polypeptide comprising the amino acid sequence of SEQ ID NO:11.
2. The isolated cDNA molecule of claim 1 , wherein the polynucleotide fragment comprises a nucleotide sequence selected from SEQ. ID NOS. 12, 14, 16, 18, 20 and 22 or encoding the amino acid sequence selected from SEQ. ID. NOS. 13, 15, 17, 19,21 and 23.
3. An isolated DNA molecule that is the complement of the cDNA molecule of claim 1 .
4. The isolated cDNA molecule of claim 1 encoding a cytochrome P450 retinoic acid metabolizing protein, comprising SEQ. ID. NO. 10 or a cDNA molecule encoding the amino acid sequence of SEQ. ID. NO. 11 or a fragment thereof, wherein said fragment retains the substrate specificity of the polypeptide comprising the amino acid sequence of SEQ ID NO:11.
5. The isolated cDNA molecule of claim 4 , wherein the retinoic acid is all trans retinoic acid(ATRA) or 9-cis-retinoic acid.
6. The isolated cDNA molecule of claim 1 , wherein the nucleotide sequence of said cDNA molecule has sequential nucleotide deletions from either the C-terminus or the N-terminus of SEQ ID NO:10.
7. A recombinant vector comprising an isolated cDNA molecule of claim 1 .
8. An isolated recombinant host cell comprising an isolated cDNA molecule of claim 1 .
9. The recombinant host cell of claim 8 wherein the isolated cDNA molecule is operatively linked to a regulatory sequence to allow expression of a peptide encoded by said cDNA molecule.
10. A diagnostic kit for identification of polymorphisms in the P450RAI-3 gene, comprising the DNA molecule that is the complement thereof, and optionally directions for the method comprising screening the P450RAI-3 gene from a human for polymorphisms, wherein detection of said polymorphisms is indicative of the occurrence of a P450RAI-3-related condition or a predisposition thereto.