5OTM

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

Summary for 5OTM

• Entry DOI: 10.2210/pdb5otm/pdb
• Descriptor: 7,8-dihydro-8-oxoguanine triphosphatase, SULFATE ION, ACETATE ION, ... (5 entities in total)
• Functional Keywords: mth1, inhibitor, complex, hydrolase, nudix, methylated nucleotide, o6-methyl-dgmp
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 2
• Total formula weight: 38057.52

Authors

Gustafsson, R.,Henriksson, L.,Jemth, A.-S.,Brautigam, L.,Carreras Puigvert, J.,Homan, E.,Warpman Berglund, U.,Helleday, T.,Stenmark, P. (deposition date: 2017-08-22, release date: 2018-09-05, Last modification date: 2024-01-17)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.8 Å)