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Vol 56(2022) N 6 p. 1003-1011; DOI 10.1134/S0026893322060024 Full Text

M.L. An1*, J.L. Miao2, 3,4,5**

Genetic and Molecular Characterization of a DASH Cryptochrome Homologous Gene from Antarctic Diatom Phaeodactylum tricornutum ICE-H

1Rizhao Polytechnic, Department of Marine Technology, Shandong, Rizhao, 276826 China
2First Institute of Oceanography, Ministry of Natural Resource, Shandong, Qingdao, 266061 China
3Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237 China
4Qingdao Key Laboratory of Marine Natural Products Research and Development, Qingdao, 266237 China
5Guangxi Academy of Sciences, Nanning, 530007 China

*rubyhawthorn@163.com
**miaojinlai@fio.org.cn
Received - 2022-04-21; Revised - 2022-07-01; Accepted - 2022-07-05

DASH-type cryptochromes (cry-DASH) belong to cryptochrome/photolyase family (CPF), that includes photolyases with the capability to repair UVB-induced DNA lesions and cryptochromes acting as photoreceptors without photorepair activity. However, recent studies demonstrated that cry-DASH can repair cyclobutane pyrimidine dimers (CPDs) in damaged single-stranded DNA (ssDNA). Up to now, cry-DASHs have been investigated in prokaryotes and eukaryotes. However, the characteristics and role of cry-DASH in Antarctic diatom were not elucidated. Here, a cry-DASH sequence (Pticry-DASH) of the Antarctic diatom Phaeodactylum tricornutum ICE-H was retrieved from the transcriptome data, amplified by RT-PCR, cloned, and then expressed in Escherichia coli. The phylogenetic analysis indicated that Pticry-DASH is most closely related to cry-DASH. The gene expression of Pticry-DASH under abiotic stress was investigated by RT-PCR. Significant transcript accumulation was revealed after exposure to UVB, blue light, high-intensity light, and high temperature. The Pticry-DASH was expressed in E. coli, purified and its activity was measured. Photo-reactivation experiments confirmed that Pticry-DASH can repair CPDs in ssDNA lesions. Taken together for the first time the cry-DASH has been identified in the Antarctic diatom. The properties and peculiarities of Pticry-DASH including UVB-induced CPDs repair function may promote the exploitation of Antarctic resources.

Phaeodactylum tricornutum, DASH cryptochrome, Pticry-DASH, DNA damage, photoreceptor



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