title: Opened-ring adducts of 5-methylcytosine and 1,5-dimethylcytosine with amines and water and evidence for an opened-ring hydrate of 2'-deoxycytidine.
abstract: A variety of nucleic acid components and related compounds undergo photoreaction with water to form so-called "photohydrates" (e.g. uracil forms 6-hydroxy-5,6-dihydrouracil). However, the corresponding hydrates of 5-methylcytosine (a minor nucleobase in eukaryotic DNA) and related compounds have not been characterized. We report the preparation of opened-ring forms of such products for 5-methylcytosine (m5C) and 1,5-dimethylcytosine (DMC). This was accomplished via thermal reaction of ring-opened amine adducts (e.g. N-carbamoyl-3-amino-2-methylacrylamidine (IVa) or N-(N'-methylcarbamoyl)-3-amino-2-methylacrylamidine (IVb)) produced by photo-induced reactions of m5C with ammonia or methylamine. When these adducts were treated with dilute trifluoroacetic acid, the amino group at the 3-position was replaced with a hydroxyl group; with IVa, N-carbamoyl-3-hydroxy-2-methylacrylamidine (Va) was formed, while reaction of IVb led to N-(N'-methylcarbamoyl)-3-hydroxy-2-methylacrylamidine (Vb). These compounds are ring-opened isomers of 5,6-dihydro-6-hydroxy-5-methylcytosine (Ia and IIa) and 5,6-dihydro-6-hydroxy-1,5-dimethylcytosine (Ib and IIb). Compounds Va and Vb each undergo thermal ring closure reactions to form two unstable compounds with chemical and UV spectral properties expected for Ia and IIa (or Ib and IIb). The latter compounds have been identified as minor products in UV-irradiated aqueous solutions of m5C and DMC. Evidence is also presented that the 2'-deoxycytidine photohydrates coexist with an opened-ring form, possibly similar in nature to Vb.
authors.affiliation: Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, CA, USA. shetlar@cgl.ucsf.edu
authors.forename: Martin D
authors.initials: MD
authors.lastname: Shetlar
authors.forename: Janet
authors.initials: J
authors.lastname: Chung
mesh_terms: D044503:5-Methylcytosine / Q000737:chemistry*; D000641:Ammonia / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000737:chemistry*; D007536:Isomerism; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D008744:Methylamines / Q000737:chemistry; D010777:Photochemistry / Q000379:methods*; D012996:Solutions; D013816:Thermodynamics; D014269:Trifluoroacetic Acid / Q000737:chemistry; D014466:Ultraviolet Rays; D014867:Water
chemical_list: D008744:Methylamines; D012996:Solutions; D014867:Water; D003841:Deoxycytidine; D044503:5-Methylcytosine; D000641:Ammonia; D004247:DNA; C027451:methylamine; D014269:Trifluoroacetic Acid