Abstract
Proteins can be methylated on the side-chain nitrogens of arginine and lysine residues or on carboxy-termini. Protein methylation is a way of subtly changing the primary sequence of a peptide so that it can encode more information. This common posttranslational modification is implicated in the regulation of a variety of processes including protein trafficking, transcription and protein-protein interactions. In this chapter, we will use the arginine methyltransferases to illustrate different approaches that have been developed to assess protein methylation. Both in vivo and in vitro methylation techniques are described, and the use of small molecule inhibitors of protein methylation will be demonstrated.
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References
Aletta, J. M., Cimato, T. R., and Ettinger, M. J. (1998) Protein methylation: a signal event in post-translational modification. Trends Biochem. Sci. 23, 89–91.
Comb, D. G., Sarkar, N., and Pinzino, C. J. (1966) The methylation of lysine residues in protein. J. Biol. Chem. 241, 1857–1862.
Paik, W. K. and Kim, S. (1968) Protein methylase I. Purification and properties of the enzyme. J. Biol. Chem. 243, 2108–2114.
Mowen, K. A., Tang, J., Zhu, W., et al. (2001) Arginine methylation of STAT1 modulates IFNalpha/beta-induced transcription. Cell 104, 731–741.
Abramovich, C., Yakobson, B., Chebath, J., and Revel, M. (1997) A protein-arginine methyltransferase binds to the intracytoplasmic domain of the IFNAR1 chain in the type I interferon receptor. EMBO J. 16, 260–266.
Strahl, B. D. and Allis, C. D. (2000) The language of covalent histone modifications. Nature 403, 41–45.
McBride, A. E. and Silver, P. A. (2001) State of the arg: protein methylation at arginine comes of age. Cell 106, 5–8.
Friesen, W. J., Massenet, S., Paushkin, S., et al. (2001) SMN, the product of the spinal muscular atrophy gene, binds preferentially to dimethylarginine-containing protein targets. Mol. Cell 7, 1111–1117.
Brahms, H., Meheus, L., de Brabandere, V., et al. (2001) Symmetrical dimethylation of arginine residues in spliceosomal Sm protein B/B and the Sm-like protein LSm4, and their interaction with the SMN protein. RNA 7, 1531–1542.
Bedford, M. T., Frankel, A., Yaffe, M. B., et al. (2000) Arginine methylation inhibits the binding of proline-rich ligands to Src homology 3, but not WW, domains. J. Biol. Chem. 275, 16,030–16,036.
Gary, J. D. and Clarke, S. (1998) RNA and protein interactions modulated by protein arginine methylation. Prog. Nucleic Acid Res. Mol. Biol. 61, 65–131.
Frankel, A., Yadav, N., Lee, J., et al. (2002) The novel human protein arginine N-methyltransferase PRMT6 is a nuclear enzyme displaying unique substrate specificity. J. Biol. Chem. 277, 3537–3543.
Lin, W. J., Gary, J. D., Yang, M. C., et al. (1996) The mammalian immediate-early TIS21 protein and the leukemia-associated BTG1 protein interact with a protein-arginine N-methyltransferase. J. Biol. Chem. 271, 15,034–15,044.
Scott, H. S., Antonarakis, S. E., Lalioti, M. D., et al. (1998) Identification and characterization of two putative human arginine methyltransferases (HRMT1L1 and HRMT1L2). Genomics 48, 330–340.
Tang, J., Gary, J. D., Clarke, S., and Herschman, H. R. (1998) PRMT 3, a type I protein arginine N-methyltransferase that differs from PRMT1 in its oligomerization, subcellular localization, substrate specificity, and regulation. J. Biol. Chem. 273, 16,935–16,945.
Chen, D., Ma, H., Hong, H., et al. (1999) Regulation of transcription by a protein methyltransferase. Science 284, 2174–2177.
Branscombe, T. L., Frankel, A., Lee, J. H., et al. (2001) Prmt5 (janus kinase-binding protein 1) catalyzes the formation of symmetric dimethylarginine residues in proteins. J. Biol. Chem. 276, 32,971–32,976.
Rea, S., Eisenhaber, F., O’Carroll, D., et al. (2000) Regulation of chromatin structure by site-specific histone H3 methyltransferases. Nature 406, 593–599.
Santos-Rosa, H., Schneider R., Bannister A. J., et al. (2002) Active genes are trimethylated at K4 of histone H3. Nature 419, 407–411.
Gary, J. D. and Clarke, S. (1995) Purification and characterization of an isoaspartyl dipeptidase from Escherichia coli. J. Biol. Chem. 270, 4076–4087.
Chevillard-Briet, M., Trouche, D., and Vandel, L. (2002) Control of CBP co-activating activity by arginine methylation. EMBO J 21, 5457–5466.
Li, H., Park S., Kilburn B., et al. (2002) Lipopolysaccharide-induced methylation of HuR, an mRNA-stabilizing protein, by CARM1. Coactivator-associated arginine methyltransferase. J. Biol. Chem. 277, 44,623–44,630.
Sommer, A., Moscatelli, D., and Rifkin, D. B. (1989) An amino-terminally extended and post-translationally modified form of a 25kD basic fibroblast growth factor. Biochem. Biophys. Res. Commun. 160, 1267–1274.
Baldwin, G. S. and Carnegie, P. R. (1971) Isolation and partial characterization of methylated arginines from the encephalitogenic basic protein of myelin. Biochem. J. 123, 69–74.
Lee, J. and Bedford, M. T. (2002) PABP1 identified as an arginine methyltransferase substrate using high-density protein arrays. EMBO Rep. 3, 268–273.
Davie, J. K. and Dent, S. Y. (2002) Transcriptional control: an activating role for arginine methylation. Curr. Biol. 12, R59–R61.
Lachner, M., O’Sullivan, R. J., and Jenuwein, T. (2003) An epigenetic road map for histone lysine methylation. J. Cell Sci. 116, 2117–2124.
Coppard, N. J., Clark, B. F., and Cramer, F. (1983) Methylation of elongation factor 1 alpha in mouse 3T3B and 3T3B/SV40 cells. FEBS Lett. 164, 330–334.
Liu, Q. and Dreyfuss, G. (1995) In vivo and in vitro arginine methylation of RNA-binding proteins. Mol. Cell Biol. 15, 2800–2808.
Frankel, A. and Clarke, S. (1999) RNase treatment of yeast and mammalian cell extracts affects in vitro substrate methylation by type I protein arginine N-methyltransferases. Biochem. Biophys. Res. Commun. 259, 391–400.
Cote, J., Boisvert, F. M., Boulanger, M. C., et al. (2003) Sam68 RNA binding protein is an in vivo substrate for protein arginine N-methyltransferase 1. Mol. Biol. Cell 14, 274–287.
Yadav, N., Lee, J., Kim, J., et al. (2003) Specific protein methylation defects and gene expression perturbations in coactivator-associated arginine methyltransferase 1-deficient mice. Proc. Natl. Acad. Sci. USA 100, 6464–6468.
Henry, M. F. and Silver, P. A. (1996) A novel methyltransferase (Hmt1p) modifies poly(A)+-RNA-binding proteins. Mol. Cell Biol. 16, 3668–3678.
Siebel, C. W. and Guthrie, C. (1996) The essential yeast RNA binding protein Np13p is methylated. Proc. Natl. Acad. Sci. USA 93, 13,641–13,646.
Bannister, A. J., Schneider, R., and Kouzarides, T. (2002) Histone methylation: dynamic or static? Cell 109, 801–806.
Najbauer, J. and Aswad, D. W. (1990) Diversity of methyl acceptor proteins in rat pheochromocytoma (PC12) cells revealed after treatment with adenosine dialdehyde. J. Biol. Chem. 265, 12,717–12,721.
Frankel, A. and Clarke, S. (2000) PRMT3 is a distinct member of the protein arginine N-methyltransferase family. Conferral of substrate specificity by a zinc-finger domain. J. Biol. Chem. 275, 32,974–32,982.
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Lee, J., Cheng, D., Bedford, M.T. (2004). Techniques in Protein Methylation. In: Dickson, R.C., Mendenhall, M.D. (eds) Signal Transduction Protocols. Methods in Molecular Biology, vol 284. Humana Press. https://doi.org/10.1385/1-59259-816-1:195
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DOI: https://doi.org/10.1385/1-59259-816-1:195
Publisher Name: Humana Press
Print ISBN: 978-1-58829-245-2
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