The Formation of Phosphate End Groups in the Radiolysis of Polynucleotides in Aqueous Solution

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The polynucleotides poly(U), poly(C), poly(A) and poly(G) have been gamma-irradiated in N2O- and N2O/O2 (4:1)-saturated aqueous solutions. Hydroxyl radicals from the radiolysis of water react with the polynucleotides thereby producing among other lesions strand breaks. Strand breakage is connected with the formation of phosphomonoester end groups. Such end groups have been determined by measuring inorganic phosphate after a three hour incubation at 37 degrees C with acid or alkaline phosphatase. In the absence of oxygen G(phosphomonoester end groups) (in units of mumol J-1) are 0.47 (poly(U)), 0.17 (poly(C)) and less than or equal to 0.04 (poly(A) and poly(G)). In the case of poly(U) and poly(C) on heating the sample for one hour at 95 degrees C prior to incubation with phosphatases the above values increased by 0.14 and 0.07 mumol J-1, resp., whereas such treatment of the purine polynucleotides still did not produce a measurable yield of phosphomonoester end groups. Comparing these values with G values for strand breakage taken from the literature, about two phosphomonoester end groups are formed per strand break in poly(U) while for poly(C) this ratio is about unity. The purine polynucleotides show very low yields of strand breakage in agreement with the negligible phosphomonoester yields. In the presence of oxygen G(phosphomonoester end groups) are 0.46 (poly(U)), 0.21 (poly(C)), and less than or equal to 0.04 (poly(A) and poly(G)). On heating, these values increase, most markedly for poly(U) and poly(C). This is possibly linked to the decomposition of unstable hydroperoxides which are formed in high yields in poly(U) and poly(C) (G = 0.7 and 0.19 mumol J-1, resp.).(ABSTRACT TRUNCATED AT 250 WORDS)