Structure analysis of Entamoeba histolytica DNMT2 (EhMeth)

TY - JOUR

T1 - Structure analysis of Entamoeba histolytica DNMT2 (EhMeth)

AU - Schulz, Eike C

AU - Roth, Heide M

AU - Ankri, Serge

AU - Ficner, Ralf

PY - 2012

Y1 - 2012

N2 - In eukaryotes, DNA methylation is an important epigenetic modification that is generally involved in gene regulation. Methyltransferases (MTases) of the DNMT2 family have been shown to have a dual substrate specificity acting on DNA as well as on three specific tRNAs (tRNA(Asp), tRNA(Val), tRNA(Gly)). Entamoeba histolytica is a major human pathogen, and expresses a single DNA MTase (EhMeth) that belongs to the DNMT2 family and shows high homology to the human enzyme as well as to the bacterial DNA MTase M.HhaI. The molecular basis for the recognition of the substrate tRNAs and discrimination of non-cognate tRNAs is unknown. Here we present the crystal structure of the cytosine-5-methyltransferase EhMeth at a resolution of 2.15 Å, in complex with its reaction product S-adenosyl-L-homocysteine, revealing all parts of a DNMT2 MTase, including the active site loop. Mobility shift assays show that in vitro the full length tRNA is required for stable complex formation with EhMeth.

AB - In eukaryotes, DNA methylation is an important epigenetic modification that is generally involved in gene regulation. Methyltransferases (MTases) of the DNMT2 family have been shown to have a dual substrate specificity acting on DNA as well as on three specific tRNAs (tRNA(Asp), tRNA(Val), tRNA(Gly)). Entamoeba histolytica is a major human pathogen, and expresses a single DNA MTase (EhMeth) that belongs to the DNMT2 family and shows high homology to the human enzyme as well as to the bacterial DNA MTase M.HhaI. The molecular basis for the recognition of the substrate tRNAs and discrimination of non-cognate tRNAs is unknown. Here we present the crystal structure of the cytosine-5-methyltransferase EhMeth at a resolution of 2.15 Å, in complex with its reaction product S-adenosyl-L-homocysteine, revealing all parts of a DNMT2 MTase, including the active site loop. Mobility shift assays show that in vitro the full length tRNA is required for stable complex formation with EhMeth.

KW - Binding Sites

KW - Catalytic Domain

KW - Crystallography, X-Ray/methods

KW - DNA (Cytosine-5-)-Methyltransferases/biosynthesis

KW - DNA Methylation

KW - Entamoeba histolytica/metabolism

KW - Humans

KW - Protein Binding

KW - Protein Conformation

KW - Protein Structure, Tertiary

KW - RNA, Transfer/chemistry

KW - S-Adenosylhomocysteine/chemistry

U2 - 10.1371/journal.pone.0038728

DO - 10.1371/journal.pone.0038728

M3 - SCORING: Journal article

C2 - 22737219

VL - 7

SP - e38728

JO - PLOS ONE

JF - PLOS ONE

SN - 1932-6203

IS - 6

ER -