Size and frequency of gaps in newly synthesized DNA of xeroderma pigmentosum human cells irradiated with ultraviolet light

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Abstract

Native newly synthesized DNA from human cells (xeroderma pigmentosum type) irradiated with ultraviolet light releases short pieces of DNA (L-DNA) when incubated with the single-strand specific S1 nuclease. This is not observed in the case of unirradiated cells. Previous experiments had shown that the L-DNA resulted from the action of S1 nuclease upon gaps, i.e., single-stranded DNA discontinuities in larger pieces of double-stranded DNA. We verified that the duplex L-DNA, that arises from the inter-gap regions upon S1 nuclease treatment, has a size which approximates the distance between two pyrimidine dimers on the same strand; this has been observed at different fluences of ultraviolet-light and indicates that the gap is related to or opposite the dimer. A method was devised to measure the size of the gaps. A Poisson distribution analysis of the percentage of the L-DNA produced as a function of S1 nuclease concentration made this possible. 65% of the gaps corresponded to stretches of 1,250 nucleotides and 35% to stretches of 150 nucleotides. These parameters have been considered in the proposition of a model for DNA synthesis on a template containing pyrimidine dimers. © 1981, The Biophysical Society. All rights reserved.

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Meneghini, R., Cordeiro-Stone, M., & Schumacher, R. I. (1981). Size and frequency of gaps in newly synthesized DNA of xeroderma pigmentosum human cells irradiated with ultraviolet light. Biophysical Journal, 33(1), 81–92. https://doi.org/10.1016/S0006-3495(81)84873-4

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