High frequency DNA rearrangements associated with mouse centromeric satellite DNA.
A DNA transformed mouse cell line, generated by the microinjection of a pBR322 plasmid containing the herpes thymidine kinase (tk) gene, was observed to exhibit a high frequency of DNA rearrangement at the site of exogenous DNA integration. The instability in this cell line does not appear to be mediated by the tk inserts or the immediately adjacent mouse DNA, but instead may be a consequence of the larger host environment at the chromosomal site of tk insertion. Results obtained from restriction analysis, in situ chromosome hybridizations, and cesium chloride density-gradient fractionations indicate that the tk inserts are organized as a single cluster of direct and inverted repeats embedded within pericentromeric satellite DNA. To determine the molecular identity of the flanking host sequences, one of the mouse-tk junction fragments was cloned, and subsequent restriction and sequence analyses revealed that this DNA fragment consists almost entirely of classical mouse satellite DNA. On the basis of these observations, we suggest that the instability in this cell line may reflect the endogenous instability or fluidity of satellite DNA.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002503:Centromere / Q000032:analysis; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004276:DNA, Satellite / Q000235:genetics; D006824:Hybridization, Genetic; D008677:Metaphase; D051379:Mice; D009693:Nucleic Acid Hybridization; D012091:Repetitive Sequences, Nucleic Acid; D013937:Thymidine Kinase; D014162:Transfection
D004276:DNA, Satellite; D013937:Thymidine Kinase; D004262:DNA Restriction Enzymes