A partial hprt gene duplication generated by non-homologous recombination in V79 Chinese hamster cells is eliminated by homologous recombination

Journal of Molecular Biology
T HelledayD Jenssen

Abstract

Here, the sequence in the hprt gene of the duplication mutant SPD8 originating from V79 Chinese hamster cells was determined. The duplication arose after non-homologous recombination between exon 6 and intron 7, resulting in an extra copy of the 3' portion of exon 6, of exon 7 and of flanking intron regions. Only a duplication of exon 7 is present in the mRNA, since the duplicated exon 6 lacks its 5' splice site and is removed during RNA processing. The findings in this study suggest that the non-homologous recombination mechanism which occurred here may have been initiated by endonucleases, rather than by a spontaneous double strand break. Subsequently, 14 spontaneous SPD8 revertants with a functional hprt gene were isolated and characterized using PCR and sequencing. The data revealed that although the SPD8 cell line arose by non-homologous recombination, it reverts spontaneously by homologous recombination. Interestingly, the downstream copy of exon 7 was restored by this process. This was indicated by the presence of a specific mutation, a T-to-G transversion, close to the breakpoint, a characteristic unique to the SPD8 clone. Our results suggest that the spontaneous reversion of this cell line by homologous recombination m...Continue Reading

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Citations

Oct 31, 2007·Molecular and Cellular Biology·Ponnari GottipatiThomas Helleday
Oct 26, 2016·International Journal of Hyperthermia : the Official Journal of European Society for Hyperthermic Oncology, North American Hyperthermia Group·Akihisa TakahashiTakeo Ohnishi
Aug 19, 2006·Human Mutation·S J WhiteJ T den Dunnen
Jan 26, 2010·Radiation Research·Jaroslaw DziegielewskiJanet E Baulch
Jul 30, 2009·Journal of Cellular Biochemistry·Anuradha KumariHua Lu
May 19, 2009·Human Genetics·Junko OshimaDaniela del Gaudio
Aug 7, 2002·Biochemical and Biophysical Research Communications·Cecilia LundinThomas Helleday
Apr 23, 2003·Journal of Molecular Biology·Cecilia LundinThomas Helleday

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