Wheat endonuclease WEN1 dependent on S-adenosyl-L-methionine and sensitive to DNA methylation status.
Ca(2+)-, Mg(2+)-dependent wheat endonuclease WEN1 with molecular mass of about 27 kDa was isolated from coleoptyles. Methylated DNA of lambda phage grown on E. coli dam(+), dcm(+) cells was hydrolyzed by WEN1 more effectively than DNA of phage grown on dam(-), dcm(-) cells. Two pH activity maxima (pH 6.5-7.5 and 9.0-10.5) were observed when double-stranded DNA was hydrolyzed. WEN1 is stable at elevated temperatures (65 degrees C ) and in wide range of pH values. WEN1 is activated by S-adenosyl-L-methionine, S-adenosyl-L-homocysteine and S-isobutyladenosine. It is a first case to show that higher eukaryote endonuclease discriminates between DNA of various methylation status and is modulated by S-AdoMet and its analogs.
Fedoreyeva
Larisa I
LI
Belozersky Institute of Physical and Chemical Biology, Lomonosov Moscow State University, Moscow, Russia.
Sobolev
Dmitriy E
DE
Vanyushin
Boris F
BF
D002413:Cations, Divalent / Q000494:pharmacology; D019175:DNA Methylation; D018744:DNA, Plant / Q000235:genetics*; D004706:Endodeoxyribonucleases / Q000302:isolation & purification / Q000378:metabolism*; D010940:Plant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000378:metabolism*; D036226:Seedlings / Q000201:enzymology; D014908:Triticum / Q000201:enzymology* / Q000235:genetics*
D002413:Cations, Divalent; D018744:DNA, Plant; D010940:Plant Proteins; D012436:S-Adenosylmethionine; D004706:Endodeoxyribonucleases; C029367:calcium magnesium dependent endodeoxyribonuclease