Regulation of gene expression by protein methylation
View Patent ↗The invention relates to the cDNA and deduced amino acid sequence of the Coactivator Associated arginine (R) Methyltransferase protein, CARM1. A method is described for the use CARM1 to regulate gene expression in vivo. CARM1 has also been used to methylate arginine residues of histones, synthetic peptides, and other proteins. A method to use CARM1 to screen for drugs that inhibit its methyltransferase activity is also described, as is a method to screen for drugs that modulate CARM1's interactions with other proteins.
1. An isolated or recombinant polypeptide comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2 and which polypeptide binds the C-terminal fragment of GRIP1.
2. The isolated or recombinant polypeptide of claim 1 wherein said peptide substantially lacks methyltransferase activity.
3. The isolated or recombinant polypeptide of claim 1 and wherein a substitution of residues 189, 190, and 191 of SEQ ID NO: 2 results in the substantial lack of methyltransferase activity.
4. The purified or recombinant polypeptide of claim 1 , wherein the polypeptide methylates histone H3.
5. An isolated or recombinant polypeptide comprising the amino acid sequence of SEQ ID NO: 3.
6. An isolated or recombinant polypeptide comprising the amino acid sequence of SEQ ID NO: 2.
7. The isolated or recombinant polypeptide of any one of claim 1 or 6 further comprising a purification domain.
8. The isolated or recombinant polypeptide of claim 7 wherein said purification domain is gluathione-S-transferase.
9. The isolated or recombinant polypeptide of any one of claim 1 or 6 further comprising a DNA binding domain.
10. The isolated or recombinant polypeptide of claim 9 wherein said DNA binding domain is the Gal4 DNA binding domain.
11. The isolated or recombinant polypeptide of any one of claim 1 or 6 further comprising a transcription activation domain.
12. The isolated or recombinant polypeptide of claim 11 wherein said transcription activation domain is the Gal4 transcription activation domain.
13. A solution comprising an isolated or recombinant polypeptide of any one of claim 1 , 3 , 5 , or 6 .