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bioRxiv · 10.1101/2022.10.26.513954

Nucleoside-driven specificity of DNA Methyltransferase

Gade, M.·Gardner, J. M.·Jain, P.·Laurino, P.
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Abstract

We have studied adenosine binding specificities of two bacterial DNA methyltransferase Taq methyltransferase (M.TaqI), and HhaI methyltransferase (M.HhaI). While these DNA methyltransferases have similar cofactor binding pocket interactions, experimental data showed different specificity for novel cofactors ((SNM) (S-guanosyl-L-methionine (SGM), S-cytidyl-L-methionine (SCM), S-uridyl-L-methionine (SUM)). Protein dynamics corroborate the experimental data on the cofactor specificities. For M.TaqI the specificity for S-adenosyl-L-methionine (SAM) is governed by the tight binding on the nucleoside part of the cofactor, while for M.HhaI the degree of freedom of the nucleoside chain allows the acceptance of other bases. The experimental data proves a catalytically productive methylation by M.HhaI binding pocket for all the SNM. Our results suggest a new route for successful design of unnatural SNM analogues for methyltransferases as a tool for cofactor engineering. Table of Content O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=82 SRC="FIGDIR/small/513954v2_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@2562aforg.highwire.dtl.DTLVardef@71de9forg.highwire.dtl.DTLVardef@1cead77org.highwire.dtl.DTLVardef@1de8682_HPS_FORMAT_FIGEXP M_FIG TOC: Methylation by DNA methyltransferase is nucleobase dependent. While M.Taq1 is specific for SAM, M.HhaI is promiscuous for other SAM analogs C_FIG

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Gade, M., Gardner, J. M., Jain, P., Laurino, P.. 2022-10-27. Nucleoside-driven specificity of DNA Methyltransferase. https://doi.org/10.1101/2022.10.26.513954

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