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Merck

A gain-of-function mouse model identifies PRMT6 as a NF-ฮบB coactivator.

Nucleic acids research (2014-06-19)
Alessandra Di Lorenzo, Yanzhong Yang, Marc Macaluso, Mark T Bedford
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Protein arginine methyltransferase 6 (PRMT6) is a nuclear enzyme that modifies histone tails. To help elucidate the biological function of PRMT6 in vivo, we generated transgenic mice that ubiquitously express PRMT6 fused to the hormone-binding portion of the estrogen receptor (ER*). The ER*-PRMT6 fusion is unstable and cytoplasmic, but upon systemic treatment with tamoxifen, it becomes stabilized and translocates into the nucleus. As a result, a dramatic increase in the H3R2me2a histone mark is observed. We found that one consequence of induced ER*-PRMT6 activation is increased IL-6 levels. IL-6 expression is regulated by the nuclear factor-kappa B (NF-ฮบB) transcription factor, and PRMT6 functions as a coactivator of this pathway. We show that PRMT6 directly interacts with RelA, and that its overexpression enhances the transcriptional activity of an ectopic NF-ฮบB reporter and endogenously regulates NF-ฮบB target genes. PRMT6 is recruited, by RelA, to selective NF-ฮบB target promoters upon TNF-ฮฑ stimulation. Moreover, ER*-PRMT6 activation causes RelA accumulation in the nucleus. In summary, we observe that PRMT6 is recruited to chromatin at selective NF-ฮบB target promoters, where it likely impacts the histone code and/or methylates other chromatin-associated proteins to facilitate transcription.

MATERIALS
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Sigma-Aldrich
Monoclonal ANTI-FLAGยฎ M2 antibody produced in mouse, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
(Z)-4-Hydroxytamoxifen, ≥98% Z isomer
Sigma-Aldrich
Anti-dimethyl-Histone H3 (Arg2) Antibody, rabbit monoclonal, Upstateยฎ, from rabbit
Sigma-Aldrich
Tamoxifen, ≥99%
Millipore
ANTI-FLAGยฎ M2 Affinity Gel, purified immunoglobulin, buffered aqueous glycerol solution