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Nucleic Acids Research, 1991, Vol. 19, No. 9 2411-2415
© 1991

MOLECULAR BIOLOGY

Strand specificity for UV-induced DNA repair and mutations in the Chinese hamster HPRT gene

Harry Vrieling1,2,*, Jaap Venema1, Marie-Louise van Rooyen1, Anneke van Hoffen1, Paola Menichini1,3, Malgorzata Z. Zdzienicka1,2, Johannes W.I.M. Simons1, Leon H.F. Mullenders1,2 and Albert A. van Zeeland1,2

1MGC, Department of Radiation Genetics and Chemical Mutagenesis, State University of Leiden Wassenaarseweg 72, 2333 AL Leiden 2J.A.Cohen Institute, Interuniversity Research Institute for Radiopathology and Radiation Protection Leiden, The Netherlands 3laboratory of Mutagenesis, National Institute for Cancer Research Viale Benedetto XV, 10, 16132 Genova, Italy

* To whom correspondence should be addressed

Received January 22, 1991. Accepted April 2, 1991.

DNA excision repair modulates the mutagenic effect of many genotoxic agents. The recently observed strand specificity for removal of UV-induced cyclobutane dinners from actively transcribed genes In mammalian cells could influence the nature and distribution of mutations In a particular gene. To investigate this, we have analyzed UV-induced DNA repair and mutagenesis in the same gene, i.e. the hypoxanthine phosphoribosyl- transferase (hprt) gene. In 23 hprt mutants from V79 Chinese hamster cells induced by 2 J/m2 UV we found a strong strand bias for mutation induction: assuming that pre-mutagenlc lesions occur at dlpyrimidine sequences, 85% of the mutations could be attributed to lesions in the nontranscrlbed strand. Analysis of DNA repair In the hprt gene revealed that more than 90% of the cyclobutane dimers were removed from the transcribed strand within 8 hours after irradiation with 10 J/m2 UV, whereas virtually no dimer removal could be detected from the nontranscribed strand even up to 24 hr after UV. These data present the first proof that strand specific repair of DNA lesions in an expressed mammalian gene is associated with a strand specificity for mutation Induction.


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