Chemical synthesis and detection of the cross-link 1-[N3-(2'-deoxycytidyl)]-2-[N1-(2'-deoxyguanosinyl)]ethane in DNA reacted with 1-(2-chloroethyl)-1-nitrosourea.

We have synthesized 1-[N3-(2'-deoxycytidyl)]-2-[N1-(2'-deoxyguanosinyl)]ethane and confirmed its structure by ultraviolet and high-resolution mass spectrometry. Treatment of calf thymus DNA with [3H](2-chloroethyl)-1-nitrosourea resulted in the formation of at least 13 DNA alkylation products that were separated by HPLC. 1-[N3-(2'-Deoxycytidyl)]-2-[N1-(2'-deoxyguanosinyl)]ethane was a minor product, accounting for 3.4% of the total DNA alkylation. The DNA cross-link 1,2-di-N7-guanylethane was formed to a similar extent (3.2%). Other minor alkylation products were O6-(2-hydroxyethyl)deoxyguanosine (1.5%) and N1-(2-hydroxyethyl)deoxyguanosine (3.8%). The principal alkylation products formed by 1-(2-chloroethyl)-1-nitrosourea (CNU) treatment of DNA were N7-(2-hydroxyethyl)guanine (36.4%), N7-(2-chloroethyl)guanine (14.6%), and phosphotriesters (26.1%). The development of analytical procedures to measure DNA alkylation products after treatment with CNU will allow us to investigate factors influencing their formation and repair.