Deciphering the DNA repair protein, Rad23 from kuruma shrimp Marsupenaeus japonicus: full-length cDNA cloning and characterization

Abstract

Aims: Lesions of DNA are removed by nucleotide excision repair (NER) process in the living systems. NER process‐related host factors are believed to aid recovery steps during viral integration. Here, we report identification and characterization of a DNA repair molecule Rad23 from kuruma shrimp Marsupenaeus japonicus. Methods and Results: The full‐length cDNA of M. japonicus Rad23 gene (MjRad23) has 1149 bp coding for a putative protein of 382 amino acids with a 5′ untranslated region (UTR) of 92 bp and 3′ UTR region of 1116 bp. Quantitative expression analysis revealed MjRad23 is constitutively expressed in all the organs of healthy shrimp, whereas with high level in muscle tissue. Although MjRad23 expression is observed in every haemolymph samplings to post‐white spot syndrome virus infection, high expression is recorded at 2 h post infection (h.p.i.). MjRad23 consists of putative functional domains including one ubiquitin domain (UBQ), two ubiquitin‐associated domains (UBA) and one heat‐shock chaperonin‐binding motif (STI1). Multiple alignment of MjRad23 with Rad23 of other species showed highly significant identity ranging from 37 to 53%; however, high homology is observed with Rad23 of Bombyx mori (BmRad23). UBQ domain region alignment revealed maximum of 66% homology with Rad23 of Apis melifera (AmRad23). MjRad23 clustered with invertebrate sector along with insect species in evolution analysis. Three‐dimensional structural analyses demonstrated the highest identity between MjRad23 and human Rad23A (hHR23A). Conclusions: The present work revealed the presence of MjRad23 gene, which is essential in DNA repair process. Further studies are required to clarify the involvement of MjRad23 in NER process. Significance and Impact of the Study: This is the first report on identification and characterization of DNA repair protein in crustaceans, which will lead to further investigation to explore the molecular mechanisms behind the NER process.

Differential expression of proteins in the gills of Litopenaeus vannamei infected with white spot syndrome virus

“…Following WSSV viral infection in shrimp, it functions as a suppressor protein. Thus, it provides the virus greater chance of viral replication and dissemination (Flegel 2007;He et al 2009;Sudhakaran et al 2011). According to Wang et al (2005), an increased expression of ubiquitin conjugating enzyme in P. vannamei (PvUbc) was detected in a time course study by real-time PCR as well as RT PCR and reported that RING H2 protein WSSV 249 from WSSV may function as E3 ligase by sequestering PvUbc for viral pathogenesis in shrimp.…”

Beyond Xeroderma Pigmentosum: DNA Damage and Repair in an Ecological Context. A Tribute to James E. Cleaver

“…Moreover, homologies between invertebrates and DNA repair proteins from a diverse array of organisms from protists to mammals have been reported ( e.g . p53, RAD23) . As an example, the protein FANCD2 associated with the human NER repair‐deficient disease, Fanconi anemia, has been found to be a paralog to sea urchin FANDCD2 .…”

BmNPV infection correlates with the enhancement of the resistance of Bombyx mori cells to UV radiation

“…Nucleotide excision repair (NER) is generally regarded to be a very important pathway for repair of UV‐induced DNA damage in both prokaryotes and eukaryotes, and generally includes damage recognition, incision, excision, repair synthesis, and ligation (Xie, Liu, Zhang, & Wang, ). Some proteins related to NER have been identified in Bombyx mori , including radiation sensitive 4 (RAD4), a key DNA repair protein that recognize various types of DNA damage (Burns, Guzder, Sung, Prakash, & Prakash, ; Chen et al, ; Gillet & Schärer, ; Jansen, Verhage, & Brouwer, ; Sinha & Häder, ; Sudhakaran et al, ; Xu, Xu, Wang, He, & Kawasaki, ). A Rad4 mutant in yeast was found to be highly sensitive to UV radiation and completely defective in NER (Zhou et al, ).…”