Crystal Structure Analysis of DNA Uridine Endonuclease Mth212 Bound to DNA

The Methanothermobacter thermautotrophicus ExoIII homologue Mth212 is a DNA uridine endonuclease

MAVLKIISWNVNGLRAVHRKGFLKWFMEEKPDILCLQEIKAAPEQLPRKLRHVEGYRSFFTPAERKGYSGVAMYTKVPPS
SLREGFGVERFDTEGRIQIADFDDFLLYNIYFPNGKMSEERLKYKLEFYDAFLEDVNRERDSGRNVIICGDFNTAHREID
LARPKENSNVSGFLPVERAWIDKFIENGYVDTFRMFNSDPGQYTWWSYRTRARERNVGWRLDYFFVNEEFKGKVKRSWIL
SDVMGSDHCPIGLEIELLEHHHHHH

reservoir solution: 10% (w/v) PEG 20000, 100mM MES-NaOH pH 6.5; protein solution: 600mM NaCl, 20mM HEPES-KOH pH 7.6, 2mM DTT; soaking in 6% (w/v) PEG 20000, 23% (v/v) glycerol, 60mM MES-NaOH pH 6.5, 200mM MnCl2, VAPOR DIFFUSION, SITTING DROP, temperature 293K

1.9 Angstrom structure of the thermophilic exonuclease III homologue Mth0212

Mth0212 in complex with a 9bp blunt end dsDNA at 1.7 Angstrom

Complex of Mth0212 and an 8bp dsDNA with distorted ends

Mth0212 in complex with a short ssDNA (CGTA)

Complex of Mth0212 and a 4 bp dsDNA with 3'-overhang

The catalytically inactive mutant Mth0212 (D151N) in complex with an 8 bp dsDNA

Mth0212 (WT) in complex with a 6bp dsDNA containing a single one nucleotide long 3'-overhang

Mth0212 (WT) crystallized in a monoclinic space group

Mth0212 in complex with ssDNA in space group P32

Mth0212 (WT) in complex with a 7bp dsDNA

The rationally designed catalytically inactive mutant Mth0212(D151N)

1.2 Angstrom structure of the exonuclease III homologue Mth0212

HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS

coordination of two manganese ions, double-strand specific 3'-5' exonuclease, AP endonuclease, 2'-deoxyuridine endonuclease, HYDROLASE

MTVLKIISWNVNGLRAVHRKGFLKWFMEEKPDILCLQEIKAAPEQLPRKLRHVEGYRSFFTPAERKGYSGVAMYTKVPPS
SLREGFGVERFDTEGRIQIADFDDFLLYNIYFPNGKMSEERLKYKLEFYDAFLEDVNRERDSGRNVIICGDFNTAHREID
LARPKENSNVSGFLPVERAWIDKFIENGYVDTFRMFNSDPGQYTWWSYRTRARERNVGWRLDYFFVNEEFKGKVKRSWIL
SDVMGSDHCPIGLEIEL