Molecular characterization and function analysis of a nucleotide excision repair gene Rad23 from Litopenaeus vannamei after Vibrio alginolyticus challenge. Issue 83 (December 2018)

• Record Type: Journal Article
• Authors: Huang, Di; Qiao, Xue-Li; Liang, Qing-jian; Wei, Wei; Kong, Jing-Rong; Huan Kang, Chang-sheng Zhao; Liu, Yuan; Wang, Wei-Na
• Is Part Of: Fish & shellfish immunology. Issue 83(2018)
• Page Start: 190
• Page End: 204
• Publication Date: 2018-12
• Subjects: Litopenaeus vannamei -- Rad23 -- Vibrio alginolyticus -- Nucleotide excision repair (NER) -- DNA damage; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2018.09.021
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: Nucleotide excision repair (NER) removes many different types of DNA lesions, and NER related host factors are reported to aid recovery steps during viral integration. Here, we report the identification and characterization of a DNA repair gene Rad23 from Litopenaeus vannamei and explore its role in innate immunity of crustaceans. Lv Rad23 contains a1149 bp open reading frame (ORF) which encodes a 382 amino acids protein with predicted theoretical isoelectric point of 4.21. Lv Rad23 was ubiquitously expressed in the muscle, eyestalk, gill, stomach, heart, legs, intestine, and hepatopancreas in order from high to low and Lv Rad23 protein was showed to be located in the cytoplasm of Drosophila S2 cells. The homology analysis showed that it has a high sequence homology with Rad23 protein from Marsupenaeus japonicus . Vibrio alginolyticus challenge induced a remarkable up-regulation of Lv Rad23 mRNA in hepatopancreas. Knocking down Lv Rad23can interfere the NER pathway by down regulating the expression of replication protein A (RPA) and proliferating cell nuclear antigen (PCNA). However it didn't cause any significant difference on total hemocyte count (THC) between Lv Rad23-silenced and non-silenced group. Lv Rad23-silenced then challenge with V. alginolyticus inducing high level of reactive oxygen species (ROS) and DNA damage in hemolymph. As well as decreased THC, which seriously diminished the innate immune system of L. vannamei . Meanwhile, the NER pathway was reactived by enhancing the expression of Lv Rad23 and promoting the production of Lv PCNA to resist apoptosis and maintain proliferation of hemolymph cells in the later stage. Our results suggest that Lv Rad23 plays a vital role in shrimp specific immune response to V. alginolytcus through its participation in NER pathway. Highlights: A crustacean Rad23 gene ( Lv Rad23) was identified in Litopenaeus vannamei. Lv Rad23 mRNA is ubiquitously expressed in all tissues with the highest level in muscle. The transcription of Lv Rad23 after V. alginplyticus challenge indicated that it is involved in pathogen infection. Knock-down of Lv Rad23 lead to severe ROS and DNA damage after V. alginplyticus challenge. Lv Rad23 was engaged in DNA repair-associated pathway.