TY - JOUR
T1 - The SOS screen in Arabidopsis
T2 - A search for functions involved in DNA metabolism
AU - Siaud, Nicolas
AU - Dubois, Emeline
AU - Massot, Sophie
AU - Richaud, Aurélien
AU - Dray, Eloïse
AU - Collier, Justine
AU - Doutriaux, Marie Pascale
PY - 2010/5/4
Y1 - 2010/5/4
N2 - 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.
AB - 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.
KW - Arabidopsis
KW - DMC1
KW - DNA metabolism
KW - RAD51
KW - RNA interference
KW - SOS
UR - http://www.scopus.com/inward/record.url?scp=77952323314&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77952323314&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2010.02.009
DO - 10.1016/j.dnarep.2010.02.009
M3 - Article
C2 - 20227352
AN - SCOPUS:77952323314
SN - 1568-7864
VL - 9
SP - 567
EP - 578
JO - DNA Repair
JF - DNA Repair
IS - 5
ER -