The genome of an organism is under constant attack from endogenous and exogenous DNA damaging factors, such as reactive radicals, radiation, and genotoxins. Therefore, DNA damage response systems to sense DNA damage, arrest cell cycle, repair DNA lesions, and/or induce programmed cell death are crucial for maintenance of genomic integrity and survival of the organism. Genome sequences revealed that, although plants possess many of the DNA damage response factors that are present in the animal systems, they are missing some of the important regulators, such as the p53 tumor suppressor. These observations suggest differences in the DNA damage response mechanisms between plants and animals. In this review the DNA damage responses in plants and animals are compared and contrasted. In addition, the function of SUPPRESSOR OF GAMMA RESPONSE 1 (SOG1), a plant-specific transcription factor that governs the robust response to DNA damage, is discussed.
Little is known about the functions of DNA polymerase λ (Pol λ) recently identified in mammals. From the genomic sequence information of rice and Arabidopsis, we found that Pol λ may be the only member of the X‐family in higher plants. We have succeeded in isolating the cDNA and recombinant protein of Pol λ in a higher plant, rice (Oryza sativa L. cv. Nipponbare) (OsPol λ). OsPol λ had activities of DNA polymerase, terminal deoxyribonucleotidyl transferase and deoxyribose phosphate lyase, a marker enzyme for base excision repair. It also interacted with rice proliferating cell nuclear antigen (OsPCNA) in a pull‐down assay. OsPCNA increased the processivity of OsPol λ. Northern blot analysis showed that the level of OsPol λ expression correlated with cell proliferation in meristematic and meiotic tissues, and was induced by DNA‐damaging treatments. These properties suggest that plant Pol λ is a DNA repair enzyme which functions in plant meristematic and meiotic tissues, and that it can substitute for Pol β and terminal deoxyribonucleotidyl transferase.
We investigated expression patterns of DNA repair genes such as the CPD photolyase, UV‐DDB1, CSB, PCNA, RPA32 and FEN‐1 genes by northern hybridization analysis and in situ hybridization using a higher plant, rice (Oryza sativa L. cv. Nipponbare). We found that all the genes tested were expressed in tissues rich in proliferating cells, but only CPD photolyase was expressed in non‐proliferating tissue such as the mature leaves and elongation zone of root. The removal of DNA damage, cyclobutane pyrimidine dimers and (6–4) photoproducts, in both mature leaves and the root apical meristem (RAM) was observed after UV irradiation under light. In the dark, DNA damage in mature leaves was not repaired efficiently, but that in the RAM was removed rapidly. Using a rice 22K custom oligo DNA microarray, we compared global gene expression patterns in the shoot apical meristem (SAM) and mature leaves. Most of the excision repair genes were more strongly expressed in SAM. These results suggested that photoreactivation is the major DNA repair pathway for the major UV‐induced damage in non‐proliferating cells, while both photoreactivation and excision repair are active in proliferating cells.
The basidiomyceteCoprinus cinereushas many advantages as a model organism for studying sexual development and meiosis, but it has been difficult to investigate using reverse-genetics methods, such as gene disruption by homologous recombination. Here, gene repression by dsRNA-mediated gene silencing was tried as an alternative method for reverse-genetics studies. It was shown that transformation of theLIM15/DMC1dsRNA expression construct (LIM15dsRNA) resulted in genomic insertion ofLIM15dsRNA and paucity of theLIM15/DMC1transcript. First,LIM15dsRNA was transformed into the homothallic strainAmutBmutto generate a homozygote in which both nuclei had a copy ofLIM15dsRNA. TheLIM15/DMC1-repressed strain showed abnormal homologous chromosome synapsis during meiosis. Basidiospore production was reduced to 16 % by the induction of dsRNA. However, approximately 60 % of basidiospores were viable. Next, a heterozygote was generated in which one nucleus had a copy ofLIM15dsRNA. The phenotype was similar to that of the homozygote. These results are not only the first demonstration of dsRNA-mediated gene silencing in a member of the homobasidiomycete fungi, to which 90 % of mushroom species belong, but also the first successful use of a reverse-genetics approach inC. cinereusresearch.