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 68.82 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.

Automated summary

The class II photolyases PiPhr1 and PiPhr2 from Antarctic diatom Phaeodactylum tricornutum ICE-H were cloned, expressed, and found to be involved in repairing DNA damaged by UV light. Their expression increased after exposure to UV radiation, blue light, and dark conditions, with in vitro assays showing their ability to repair CPDs using blue light. This discovery contributes to understanding the functional role of these genes in Antarctic diatoms.