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Content archived on 2024-04-15

MOLECULAR BASIS OF RADIATION-INDUCED MUTAGENESIS FROM BACTERIA TO HUMANS. NEW EXPERIMENTAL SYSTEMS

Objective


A new experimental system for examining the molecular basis of radiation induced mutagenesis, from bacteria to humans, has been developed and an error prone SOS inducible deoxyribonucleic acid (DNA) polymerase, capable of copying typical noncoding DNA damage, has been identified. This damage is in the abasic site which is known to be produced by ionising radiation, both directly (depurination) and following the activity of repair enzymes (N-glycosylases). The inducible mutagenic polymerase was found to be Escherichia coli DNA polymerase II.

A new experimental system is being developed for examining the molecular basis of radiation induced mutagenesis in species ranging from bacteria to humans.

In a three years long collaborative effort with the laboratory of Dr M Goodman (Univ of Southern California) we have succeeded in identifying an error-prone SOS-inducible DNA polymerase capable of copying a typical noncoding DNA damage. This damage is in the abasic site which is known to be produced by ionising radiation both directly (depurination) and following the activity of repair enzymes (N-glycosylases) removing the damaged base (eg thymine glycol) which was incorporated into a specific site of a synthetic oligonucleotide substrate. The inducible mutagenic polymerase is the E. coli DNA polymerase II (2).
THE FOLLOWING RESEARCH IS PLANNED FOR THE PERIOD TO 1989:

1985-1986: ISOLATION OF DNA MISMATCH RECOGNIZING PROTEINS FROM E.COLI ANTIBODIES RAISED AGAINST HYBRID PROTEINS "MISMATCH REPAIR/BETA-GALACTOSIDASE" WILL BE USED TO PURIFY THE NATIVE MISMATCH REPAIR COMPLEX FROM E.COLI.

1986-1987: CONDITIONS FOR DETECTION OF SINGLE BASE SUBSTITUTION OR FRANESHIFT MUTATIONS IN DEFINED HUMAN GENES.

1987-1988: SPONTANEOUS AND INDUCED MUTAGENESIS OF THE SHUTTLE VECTORS/MUTATION PROBES IN E.COLI YEAST, PLANT AND MAMMALIAN CELLS.

1988-1989: SEQUENCING OF LAC- MUTATIONS PRODUCED IN E.COLI YEAST, PLANT, AND MAMMALIAN CELLS IN THE UNIVERSAL GENETIC PROBE.

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ASSOCIATION POUR LE DEVELOPPEMENT DE LA RECHERCHE EN BIOLOGIE MOLECULAIRE
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INSTITUT JACQUES MONOD 2 PLACE JUSSIEU
75251 PARIS
France

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