Expression pattern analysis of DNA repair-related and DNA damage response genes revealed by 55K oligomicroarray upon UV-B irradiation in the intertidal copepod, Tigriopus japonicus

Abstract

Ultraviolet-B (UV-B) radiation affects the genome stability of aquatic organisms by absorption of certain wavelength at the molecular level. Recently, extensive gene information has been identified from the intertidal copepod, Tigriopus japonicus. Here, we developed a 55K (54,254 genes) oligomicroarray and tested its usefulness to identify the effect of single dose of UV-B irradiation (12 kJ/m(2)) on transcriptomes of the copepod T. japonicus. A total of 35,361 spots were identified to be significantly modulated on the 55K oligomicroarray by hierarchical clustering after exposure to UV-B irradiation over 48 h (6, 12, 24, and 48 h). Of them, 1300 and 588 genes were observed to be up-regulated and down-regulated at all time points, respectively. Particularly, it was observed that several genes involved in DNA repair mechanism were significantly modulated in the UV-B-exposed T. japonicus by microarray and quantitative real-time RT-PCR analysis. In detail, UV-B irradiation specifically up-regulated some genes in non-homologous end-joining (NHEJ), homologous recombination (HR), base excision repair (BER), and mismatch repair (MMR) pathways. On the other hand, a majority of down-regulated genes were representatives for the nucleotide excision repair (NER) mechanism. These results demonstrated that DNA damage would be induced by UV-B irradiation in this species, resulting in reliable induction or repression of various DNA repair mechanism on UV-B-induced DNA damage. In this report, we suggest that a high density microarray-based approach for risk assessment of UV-B irradiation would be useful to elucidate the mechanistic analysis in a non-model organism. This study could also provide a better understanding of molecular mechanisms of cellular protection against UV-B-induced stress.