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Nucleic Acids Research, 1992, Vol. 20, No. 16 4269-4274
© 1992

MOLECULAR BIOLOGY

Molecular analysis of DNA junctions produced by illegitimate recombination in human cells

Anne Stary and Alain Sarasin

Laboraory of Molecular Genetics, Insitute de Recherches Scientifiques sur le Cancer BP no.8, 84801 Villejuif, France

Received April 30, 1992. Revised June 30, 1992. Accepted June 30, 1992.

In a human HeLa derived-cell line carrying permanently a single integrated copy of an SV40 shuttle vector, the transient expression of the SV40 T-antlgen led to the production of heterogeneous populations of circular DNA molecules which retained both integrated vector and its surrounding cellular sequences. Comparison between the integrated copy and the linear maps of 80 different plasmids rescued in bacteria suggested that the formation of circular DNA was the result of bidirectionnal replication from the SV40 origin of replication followed by a single intramolecular joining leading to the cyclization of the replicated molecules. Sequence analysis of 45 recomblnational junctions demonstrated that the cyclization occurred via illegitimate recombination process which did not require preferential nucleotide sequence at the joining sites. However, extensive characterization of recombination junctions revealed that the sequences Involved in the recombination at each side of the SV40 origin of replication were not randomly distributed, suggesting the presence of regions which were more prone to be involved in the illegitimate recombination process in human cells. Search of common features usually Implied in illegitimate recombination in mammalian cells revealed some association of these regions with palindromes, A + T-rich DNA segments, alternating purine/pyrimldlne sequences and Alu family repeats.


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