5OTM

Crystal structure of human MTH1 in complex with O6-methyl-dGMP

5OTM の概要

• エントリーDOI: 10.2210/pdb5otm/pdb
• 分子名称: 7,8-dihydro-8-oxoguanine triphosphatase, SULFATE ION, ACETATE ION, ... (5 entities in total)
• 機能のキーワード: mth1, inhibitor, complex, hydrolase, nudix, methylated nucleotide, o6-methyl-dgmp
• 由来する生物種: Homo sapiens (Human)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 38057.52

構造登録者

Gustafsson, R.,Henriksson, L.,Jemth, A.-S.,Brautigam, L.,Carreras Puigvert, J.,Homan, E.,Warpman Berglund, U.,Helleday, T.,Stenmark, P. (登録日: 2017-08-22, 公開日: 2018-09-05, 最終更新日: 2024-01-17)

主引用文献

Jemth, A.S.,Gustafsson, R.,Brautigam, L.,Henriksson, L.,Vallin, K.S.A.,Sarno, A.,Almlof, I.,Homan, E.,Rasti, A.,Warpman Berglund, U.,Stenmark, P.,Helleday, T.
MutT homologue 1 (MTH1) catalyzes the hydrolysis of mutagenic O6-methyl-dGTP.
Nucleic Acids Res., 46:10888-10904, 2018

PubMed Abstract: Nucleotides in the free pool are more susceptible to nonenzymatic methylation than those protected in the DNA double helix. Methylated nucleotides like O6-methyl-dGTP can be mutagenic and toxic if incorporated into DNA. Removal of methylated nucleotides from the nucleotide pool may therefore be important to maintain genome integrity. We show that MutT homologue 1 (MTH1) efficiently catalyzes the hydrolysis of O6-methyl-dGTP with a catalytic efficiency similar to that for 8-oxo-dGTP. O6-methyl-dGTP activity is exclusive to MTH1 among human NUDIX proteins and conserved through evolution but not found in bacterial MutT. We present a high resolution crystal structure of human and zebrafish MTH1 in complex with O6-methyl-dGMP. By microinjecting fertilized zebrafish eggs with O6-methyl-dGTP and inhibiting MTH1 we demonstrate that survival is dependent on active MTH1 in vivo. O6-methyl-dG levels are higher in DNA extracted from zebrafish embryos microinjected with O6-methyl-dGTP and inhibition of O6-methylguanine-DNA methyl transferase (MGMT) increases the toxicity of O6-methyl-dGTP demonstrating that O6-methyl-dGTP is incorporated into DNA. MTH1 deficiency sensitizes human cells to the alkylating agent Temozolomide, a sensitization that is more pronounced upon MGMT inhibition. These results expand the cellular MTH1 function and suggests MTH1 also is important for removal of methylated nucleotides from the nucleotide pool.

PubMed: 30304478
DOI: 10.1093/nar/gky896

実験手法

X-RAY DIFFRACTION (1.8 Å)