Two class II CPD photolyases, PiPhr1 and PiPhr2, with CPD repair activity from the Antarctic diatom Phaeodactylum tricornutum ICE-H

Two gene of class II photolyases, PiPhr1 (1833 bp) and PiPhr2 (1809 bp), from the Antarctic diatom Phaeodactylum tricornutum ICE-H were cloned, the recombinant proteins expressed and purified. The molecular weight of the recombinant photolyases were determined to be 68 kDa with a pI of 9.04 and 68.8...

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Bibliographic Details
Published in3 Biotech Vol. 11; no. 8; p. 377
Main Authors An, Meiling, Qu, Changfeng, Miao, Jinlai, Sha, Zhenxia
Format Journal Article
LanguageEnglish
Published Cham Springer International Publishing 01.08.2021
Springer Nature B.V
Subjects
Agriculture
Bioinformatics
Biomaterials
Biotechnology
Cancer Research
Chemistry
Chemistry and Materials Science
Deoxyribonucleic acid
DNA
DNA repair
Marine microorganisms
Molecular weight
Original
Original Article
Phaeodactylum tricornutum
Photolyase
Photoreactivation
Repair
Stem Cells
Ultraviolet radiation
CPD photolyase
Photoreactivation
Antarctic diatom
ICE-H
CPD
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Summary:Two gene of class II photolyases, PiPhr1 (1833 bp) and PiPhr2 (1809 bp), from the Antarctic diatom Phaeodactylum tricornutum ICE-H were cloned, the recombinant proteins expressed and purified. The molecular weight of the recombinant photolyases were determined to be 68 kDa with a pI of 9.04 and 68.82 with a pI of 7.31, respectively. Activity studies showed that both the recombinant enzymes were involved in the repair DNA damaged by UV light, that is they were most likely photolyases involved in photorepair of DNA. Further confirmation of this function was demonstrated by the increased expression of PiPhr1 and PiPhr2 after exposure to UV radiation, blue light and dark conditions by qRT-PCR. In summary, PiPhr1 and PiPhr2 were up regulated by UVB irradiation and blue light at 0.5 h and 3 h. Longtime (3 h) exposure to dark also increased the expression of PiPhr1 and PiPhr2 . In vitro photoreactivation assays showed that PiPhr1 and PiPhr2 could repair CPDs utilizing blue light. This is the first time CPD Class II photolyase has been reported from Antarctic diatom. These results will add to the knowledge of the diatom CPF family and assist in understanding the functional role of these genes in Antarctic diatoms.
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ISSN:2190-572X
2190-5738
DOI:10.1007/s13205-021-02927-0