Prevention of 1-(3-deoxycytidyl),2-(1-deoxyguanosinyl)ethane cross-link formation in DNA by rat liver O6-alkylguanine-DNA alkyltransferase.
Previous studies have shown that the haloethylnitrosoureas introduce the cross-link 1-(3-deoxycytidyl),2-(1-deoxyguanosinyl)ethane into DNA. This structure is evidently formed by the following sequence of events: an initial attack of a haloethyl group on the O6 position of guanine, formation of the reactive intermediate, 1,O6-ethanoguanine, and reaction of this intermediate with deoxycytidine in the opposite DNA strand. To investigate the role of O6-alkylguanine-DNA alkyltransferase in preventing the formation of this cross-link, a DNA substrate containing O6-fluoroethylguanine has been prepared by reacting DNA with N-2-fluoroethyl-N'-cyclohexyl-N-nitrosourea. The O6-fluoroethylguanine content of this substrate decreases when it is incubated at 37 degrees C and pH 7.8 in the absence of repair factors because of the chemical instability of O6-fluoroethylguanine; however, this loss is accelerated by the addition of rat liver O6-alkylguanine-DNA alkyltransferase, indicating that this repair factor recognizes and repairs O6-fluoroethylguanine in DNA; furthermore, by using [chloroethyl-14C]N-chloroethyl-N'-cyclohexyl-N-nitrosourea, it can be shown directly that the addition of rat liver O6-alkylguanine-DNA alkyltransferase prevents 1-(3-deoxycytidyl),2-(1-deoxyguanosinyl)ethane formation. These studies link the presence of repair activity to the prevention of a specific cytotoxic lesion in DNA.
Ludlum
D B
DB
Mehta
J R
JR
Tong
W P
WP
D000478:Alkylation; D003432:Cross-Linking Reagents; D004247:DNA; D004260:DNA Repair / Q000187:drug effects; D066298:In Vitro Techniques; D007700:Kinetics; D008130:Lomustine / Q000031:analogs & derivatives; D008780:Methyltransferases / Q000378:metabolism*; D019853:O(6)-Methylguanine-DNA Methyltransferase
D003432:Cross-Linking Reagents; C017318:1-(2-fluoroethyl)-3-cyclohexyl-1-nitrosourea; D008130:Lomustine; D004247:DNA; D008780:Methyltransferases; D019853:O(6)-Methylguanine-DNA Methyltransferase