Transposition of an Alu-containing element induced by DNA-advanced glycosylation endproducts.
Advanced glycosylation endproducts react with DNA and cause mutations and DNA transposition in bacteria. To investigate the mutagenic effect of advanced glycosylation in mammalian cells, plasmid DNA containing the lacI mutagenesis marker was modified by advanced glycosylation endproducts in vitro, transfected into murine lymphoid cells, recovered, and analyzed for mutations, plasmid size changes, and the presence of shared insertion sequences. An 853-bp host-derived DNA sequence, designated INS-1, was identified as an insertion element common to plasmids recovered from multiple independent transfections. Modification of DNA by advanced glycosylation increased by 60-fold the apparent frequency of INS-1 transposition: from 0.025% to 1.5%. The INS-1 element contains a 180-bp region that is homologous to the Alu repetitive sequence family. INS-1 was also observed to be present within larger insertional mutations and, in two cases, an apparently truncated version of INS-1 that lacks the Alu region was identified. These results demonstrate the experimental induction of DNA transposition involving mammalian chromosomal elements and suggest that advanced glycosylation may play a role in the formation of Alu-containing insertions that have been found to disrupt human genes.
Picower Institute for Medical Research, Manhasset, NY 11030.
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Bucala
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Lee
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Rourke
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Cerami
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics*; D005819:Genetic Markers; D017127:Glycation End Products, Advanced / Q000494:pharmacology*; D051379:Mice; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009153:Mutagens; D009838:Oligodeoxyribonucleotides; D010957:Plasmids / Q000187:drug effects*; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping; D014162:Transfection
D004251:DNA Transposable Elements; D005819:Genetic Markers; D017127:Glycation End Products, Advanced; D009153:Mutagens; D009838:Oligodeoxyribonucleotides; D004247:DNA