Nucleic Acids Research, 1988, Vol. 16, No. 23 11319-11326
© 1988
MOLECULAR BIOLOGY |
ADP-ribosylation is involved in the integration of foreign DNA into the mammalian cell genome
Cell and Molecular Biology Laboratory, Biology Building, University of Sussex Brighton, BNI 9QG, UK 1Molecular Genetics Unit, Department of Obstetrics and Gynaecology, King's College School of Medicine and Dentistry Denmark Hill, London, SE5 8RX, UK
Received October 13, 1988. Accepted November 4, 1988.
The most commonly used DNA transfection method, which employs the calcium phosphate co-precipitation of the donor DNA, involves several discrete steps (1,2). These include the uptake of the donor DNA by the recipient cells, the transport of the DNA to the nucleus, transient expression prior to integration into the host cell genome, concatenation and integration of the transfected DNA into the host cell genome and finally the stable expression of the integrated genes (2,3). Both the concatenation and the integration of the donor DNA into the host genome involve the formation and ligation of DNA strand-breaks. In the present study we demonstrate that the nuclear enzyme, adenosine diphosphoribosyl transferase (ADPRT, E.C. 2.4.2.30 [EC] ), which is dependent on the presence of DNA strand breaks for its activity (4,5) and necessary for the efficient ligation of DNA strand-breaks In eukaryotic cells (4,6), is required for the integration of donor DNA into the host genome. However, ADPRT activity does not influence the uptake of DNA into the cell, its episomal maintenance or replication, nor its expression either before or after integration into the host genome. These observations strongly suggest the involvement of ADPRT activity in eukaryotic DNA recombination events.
An abstract of this work was presented at the 8th International Symposium on ADP-bosylation, Fortworth, Texas, May 30th to June 3rd 1987
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