Identification of radiation-responsive genes in vitro using a gene trap strategy predicts for modulation of expression by radiation in vivo.
A large number of genes are known to be responsive to ionizing radiation, and there is strong evidence for the existence of inducible radiation resistance in mammalian cells. We have developed a gene trap insertional mutagenesis strategy to identify novel genes involved in responses to radiation. Using this approach, we have isolated four gene-trap integrations in embryonic stem cells. In three cases (9A, 3E and 9H) the trapped genes are radiation-inducible, and in one (7D) the gene is down-regulated. Sequence analysis of fusion transcripts from three of the integrations indicate one novel gene (3E), the mouse homologue (9A) of a known but uncharacterized human gene that encodes a protein with significant homology to several GTPase-activating proteins and a murine locus, Mym (9H). The embryonic stem cell clone with the 9A insertion was introduced into the mouse germline, and the in vivo expression pattern of 9A was studied in detail. A unique, spatially restricted pattern of expression in embryos and adult animals was observed. There is tissue-specific in vivo induction of the 9A gene in adult mice by radiation. This study demonstrates the potential of the gene trap approach for the identification and functional analysis of novel radiation-regulated genes. Similar strategies may facilitate the discovery and characterization of genes involved in other cellular stress responses.
Vallis
Katherine A
KA
Department of Radiation Oncology, University of Toronto, Ontario M5G 2M9, Canada. katherine.vallis@rmp.uhn.on.ca
Chen
Zhuo
Z
Stanford
William L
WL
Yu
Mei
M
Hill
Richard P
RP
Bernstein
Alan
A
D000595:Amino Acid Sequence; D000818:Animals; D002678:Chimera; D002999:Clone Cells / Q000528:radiation effects; D018920:Coculture Techniques; D004249:DNA Damage; D004624:Embryo Transfer; D004622:Embryo, Mammalian / Q000166:cytology; D020224:Expressed Sequence Tags; D005260:Female; D005326:Fetal Proteins / Q000235:genetics; D005786:Gene Expression Regulation / Q000528:radiation effects*; D005796:Genes; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D018095:Germ-Line Mutation; D006801:Humans; D007438:Introns / Q000235:genetics; D007763:Lac Operon; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013234:Stem Cells / Q000528:radiation effects; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D005326:Fetal Proteins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase