The SOS screen in Arabidopsis: a search for functions involved in DNA metabolism.
The SOS screen, as originally described by Perkins et al. (1999) [7], was setup with the aim of identifying Arabidopsis functions that might potentially be involved in the DNA metabolism. Such functions, when expressed in bacteria, are prone to disturb replication and thus trigger the SOS response. Consistently, expression of AtRAD51 and AtDMC1 induced the SOS response in bacteria, even affecting E. coli viability. 100 SOS-inducing cDNAs were isolated from a cDNA library constructed from an Arabidopsis cell suspension that was found to highly express meiotic genes. A large proportion of these SOS(+) candidates are clearly related to the DNA metabolism, others could be involved in the RNA metabolism, while the remaining cDNAs encode either totally unknown proteins or proteins that were considered as irrelevant. Seven SOS(+) candidate genes are induced following gamma irradiation. The in planta function of several of the SOS-inducing clones was investigated using T-DNA insertional mutants or RNA interference. Only one SOS(+) candidate, among those examined, exhibited a defined phenotype: silenced plants for DUT1 were sensitive to 5-fluoro-uracil (5FU), as is the case of the leaky dut-1 mutant in E. coli that are affected in dUTPase activity. dUTPase is essential to prevent uracil incorporation in the course of DNA replication.
Institut de Biologie des Plantes, CNRS UMR8618, Bâtiment 630, Université Paris Sud 11, 91405 Orsay Cedex, France. nicolas.siaud@u-psud.fr
Nicolas
N
Siaud
Emeline
E
Dubois
Sophie
S
Massot
Aurélien
A
Richaud
Eloïse
E
Dray
Justine
J
Collier
Marie-Pascale
MP
Doutriaux
D000818:Animals; D017360:Arabidopsis / Q000166:cytology / Q000235:genetics* / Q000378:metabolism*; D002478:Cells, Cultured; D004269:DNA, Bacterial / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology; D018506:Gene Expression Regulation, Plant; D006801:Humans; D008540:Meiosis / Q000235:genetics; D051379:Mice; D050296:Microbial Viability; D016254:Mutagenesis, Insertional; D020411:Oligonucleotide Array Sequence Analysis; D010940:Plant Proteins / Q000235:genetics; D011755:Pyrophosphatases / Q000172:deficiency / Q000235:genetics; D034622:RNA Interference; D018749:RNA, Plant / Q000378:metabolism; D051135:Rad51 Recombinase / Q000235:genetics; D013014:SOS Response, Genetics; D012689:Sequence Homology, Nucleic Acid
D004269:DNA, Bacterial; D018076:DNA, Complementary; D018744:DNA, Plant; D010940:Plant Proteins; D018749:RNA, Plant; C036483:T-DNA; D051135:Rad51 Recombinase; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase