Crystal structure of the methyltransferase subunit of human mitochondrial Ribonuclease P (MRPP1) bound to S-adenosyl-methionine (SAM)

Abstract DNA methylation, which requires the universal methyl donor S ‐adenosyl‐L‐methionine (SAM), plays a pivotal role in eukaryotic gene regulation and when dysregulated, can result in severe alterations in cellular function. An emerging approach to further understand DNA methylation utilizes azide‐ and alkyne‐functionalized N ‐mustard SAM analogues as biochemical tools to probe sites of DNA methylation. While the successful utility of these substituted analogues has been demonstrated with prokaryotic DNA methyltransferases, their utility with physiologically‐relevant eukaryotic DNA methyltransferase 1 (DNMT1) is examined for the first time here. A fluorescence‐based magnetic bead assay was validated in initial experiments to measure the extent of DNA modification by the N ‐mustard analogues using Spiroplasma methylase, M.SssI, a prokaryotic model of DNMT1. Subsequent analysis with DNMT1 revealed limited utility of the analogues, as added azide‐ and alkyne‐functionality appears to directly impact binding to DNMT1.

DNA methyltransferase

DNMT1

10.1002/slct.201902940

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Protein methylation in animal cells II. Inhibition of N-methyltransferase by analogs of

Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis (1978)

Pierre Casellas Philippe Jeanteur

Ribose

Side chain

Moiety

10.1016/0005-2787(78)90078-3

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Crystal structure of dengue virus methyltransferase without S-adenosyl-L-methionine

Antiviral Research (2014)

Christian G. Noble Shihua Li Hongping Dong S. Chew Pei‐Yong Shi

Transferase

O-methyltransferase

10.1016/j.antiviral.2014.09.003

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Citations (22)

Structure of Protein Phosphatase Methyltransferase 1 (PPM1), a Leucine Carboxyl Methyltransferase Involved in the Regulation of Protein Phosphatase 2A Activity

Journal of Biological Chemistry (2004)

Nicolas Leulliot Sophie Quevillon‐Chéruel Isabelle Sorel I. Li de la Sierra-Gallay Bruno Collinet

The important role of the serine/threonine protein phosphatase 2A (PP2A) in various cellular processes requires a precise and dynamic regulation of PP2A activity, localization, and substrate specificity. The regulation of the function of PP2A involves the reversible methylation of the COOH group of the C-terminal leucine of the catalytic subunit, which, in turn, controls the enzyme's heteromultimeric composition and confers different protein recognition and substrate specificity. We have determined the structure of PPM1, the yeast methyltransferase responsible for methylation of PP2A. The structure of PPM1 reveals a common S-adenosyl-l-methionine-dependent methyltransferase fold, with several insertions conferring the specific function and substrate recognition. The complexes with the S-adenosyl-l-methionine methyl donor and the S-adenosyl-l-homocysteine product and inhibitor unambiguously revealed the co-substrate binding site and provided a convincing hypothesis for the PP2A C-terminal peptide binding site. The structure of PPM1 in a second crystal form provides clues to the dynamic nature of the PPM1/PP2A interaction.

DUSP6

Protein phosphatase 1

10.1074/jbc.m311484200

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Citations (85)

Enzymatic stereospecific preparation of fluorescent S-adenosyl-l-methionine analogs

Analytical Biochemistry (2009)

Otmar M. Ottink Frank H. T. Nelissen Yvonne H. W. Derks Sybren S. Wijmenga Hans A. Heus

Stereospecificity

10.1016/j.ab.2009.09.013

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Citations (30)