mAM facilitates conversion by ESET of dimethyl to trimethyl lysine 9 of histone H3 to cause transcriptional repression Journal Article
| Authors: | Wang, H.; An, W.; Cao, R.; Xia, L.; Erdjument-Bromage, H.; Chatton, B.; Tempst, P.; Roeder, R. G.; Zhang, Y. |
| Article Title: | mAM facilitates conversion by ESET of dimethyl to trimethyl lysine 9 of histone H3 to cause transcriptional repression |
| Abstract: | Methylation of histone tails plays an important role in chromatin structure and function. Previously, we reported that ESET/SETDB1 is a histone methyltransferase (HMTase). Here, we show that SETDB1 tightly associates with the human homolog of mAM, a murine ATFa-associated factor. Although recombinant ESET can methylate lysine 9 of histone H3 (H3-K9), its activity is severely compromised when compared to that of the ESET/mAM complex. mAM stimulates ESET enzymatic activity by increasing the Vmax and decreasing the K m. Importantly, mAM facilitates the ESET-dependent conversion of dimethyl H3-K9 to the trimethyl state both in vitro and in vivo. Chromatin-based transcription and ChIP analyses demonstrate that mAM enhances ESET-mediated transcriptional repression in a SAM-dependent manner, and this repression correlates with H3-K9 trimethylation at the promoter. Thus, our studies establish that promoter H3-K9 trimethylation is the cause of transcriptional repression and that mAM/hAM facilitates conversion of H3-K9 dimethyl to trimethyl by ESET/SETDB1. |
| Keywords: | protein expression; methylation; promoter region; nonhuman; polymerase chain reaction; protein analysis; animals; mice; cell line; protein binding; rna interference; genetic transcription; in vivo study; transcription, genetic; in vitro study; dose-response relationship, drug; hela cells; time factors; transcription factors; blotting, western; correlation analysis; methyltransferase; methyltransferases; transcription regulation; amino acid sequence; molecular sequence data; genetic transfection; kinetics; protein purification; chromatin; histone-lysine n-methyltransferase; histone h3; recombinant proteins; substrate specificity; western blotting; murinae; binding sites; enzyme specificity; enzyme substrate complex; repressor proteins; histones; enzymes; chemical reaction; lysine; electrophoresis, polyacrylamide gel; velocity; enzyme active site; promoter regions (genetics); silver staining; precipitin tests; humans; article |
| Journal Title: | Molecular Cell |
| Volume: | 12 |
| Issue: | 2 |
| ISSN: | 1097-2765 |
| Publisher: | Cell Press |
| Date Published: | 2003-08-01 |
| Start Page: | 475 |
| End Page: | 487 |
| Language: | English |
| DOI: | 10.1016/j.molcel.2003.08.007 |
| PUBMED: | 14536086 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Export Date: 12 September 2014 -- Source: Scopus |
