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  • Title: Analysis of mutations induced by 4'-hydroxymethyl-4,5',8-trimethylpsoralen and UVA in Escherichia coli lac Z gene and its regulatory region.
    Author: Collet M, Piette J.
    Journal: J Photochem Photobiol B; 1993 Oct; 20(2-3):145-52. PubMed ID: 8271115.
    Abstract:
    The mutagenic consequences of covalent adducts induced in M13mp19 DNA by 4'-hydroxymethyl-4,5',8-trimethylpsoralen (HMT) and UVA have been determined in a forward mutational system capable of detecting all classes of mutagenic events. The photoreaction mediated by HMT has been carried out at a very low molar ratio of HMT to DNA which favours the induction of cross-links between high affinity reaction sites. When damaged M13mp19 DNA is used to transfect competent Escherichia coli K12 JM105 cells, a five-fold increase in mutation frequency is observed at 3.5% survivors when measured as a loss of beta-galactosidase alpha-complementation. The enhanced mutation frequency is largely due to base substitutions, frameshift events and large deletions. The single nucleotide substitutions occur both in the lac Z coding sequence and in its regulatory region. Transversion and transition have been detected with a predominant form consisting of A.T-to-G.C transversion at position +159. Frameshift mutations have been observed at five positions while three large deletions removing either part of the coding sequence or both the coding and the regulatory regions have been detected with a higher frequency. The spectrum of base substitutions detected between the M13 lac Z- phages surviving to the treatment is totally different from those appearing spontaneously whereas several frameshift events or deletions can already be detected between the spontaneous mutations. Despite the presence of these spontaneous hot spots, the spectrum of mutations recovered after HMT photoaddition appears to be unique and a detailed analysis of the different classes of mutations indicates an important role of cross-links in the production of mutations.
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