Molecular cloning, sequence characterization, and expression analysis of C-type lectin (CTL) and ER-Golgi intermediate compartment 53-kDa protein (ERGIC-53) homologs from the freshwater prawn, Macrobrachium rosenbergii

Snigdha Baliarsingh; Sonalina Sahoo; Yong Hun Jo; Yeon Soo Han; Arup Sarkar; Yong Seok Lee; Jyotirmaya Mohanty; Bharat Bhusan Patnaik

doi:10.1007/s10499-022-00845-3

Molecular cloning, sequence characterization, and expression analysis of C-type lectin (CTL) and ER-Golgi intermediate compartment 53-kDa protein (ERGIC-53) homologs from the freshwater prawn, Macrobrachium rosenbergii

Aquac Int. 2022;30(2):1011-1035. doi: 10.1007/s10499-022-00845-3. Epub 2022 Feb 5.

Authors

Snigdha Baliarsingh¹, Sonalina Sahoo², Yong Hun Jo³, Yeon Soo Han³, Arup Sarkar⁴, Yong Seok Lee⁵, Jyotirmaya Mohanty², Bharat Bhusan Patnaik¹

Affiliations

¹ PG Department of Biosciences and Biotechnology, Fakir Mohan University, Vyasa Vihar, Nuapadhi, Balasore, 756089 Odisha India.
² Fish Genetics and Biotechnology Division, ICAR-Central Institute of Freshwater Aquaculture, Kausalyaganga, Bhubaneswar, 751002 Odisha India.
³ Department of Applied Biology, Institute of Environmentally-Friendly Agriculture, School of Agriculture and Life Sciences, Chonnam National University, Gwangju, South Korea.
⁴ School of Biotech Sciences, Trident Academy of Creative Technology, Chandaka Industrial Estate, Chandrasekharpur, Bhubaneswar, 751024 Odisha India.
⁵ School of Life Sciences and Biotechnology, College of Natural Sciences, Soonchunhyang University, Asan City, Asan, South Korea.

Abstract

Lectin protein families are diverse and multi-functional in crustaceans. The carbohydrate-binding domains (CRDs) of lectins recognize the molecular patterns associated with pathogens and orchestrate important roles in crustacean defense. In this study, two lectin homologs, a single CRD containing C-type lectin (CTL) and an L-type lectin (LTL) domain containing endoplasmic reticulum Golgi intermediate compartment 53 kDa protein (ERGIC-53) were identified from the freshwater prawn, Macrobrachium rosenbergii. The open reading frames of MrCTL and MrERGIC-53 were 654 and 1,515 bp, encoding polypeptides of 217 and 504 amino acids, respectively. Further, MrCTL showed a 20-amino acid transmembrane helix region and 10 carbohydrate-binding residues within the CRD. MrERGIC-53 showed a signal peptide region, a type-I transmembrane region, and a coiled-coil region at the C-terminus. Phylogenetic analysis revealed a close relationship between MrCTL and MrLectin and M. nipponense CTL (MnCTL), whereas MrERGIC-53 shared high sequence identity with Eriocheir sinensis ERGIC-53 and Penaeus vannamei MBL-1. A homology-based model predicted small carbohydrate-combining sites with a metal-binding site for ligand binding (Ca²⁺ binding site) in MrCTL and beta-sheets connected by short loops and beta-bends forming a dome-shaped beta-barrel structure representing the LTL domain of MrERGIC-53. Quantitative real-time polymerase chain reaction detected MrCTL and MrERGIC-53 transcripts in all examined tissues, with particularly high levels observed in hemocytes, hepatopancreas, and mucosal-associated tissues, such as the stomach and intestine. Further, the expression levels of MrCTL and MrERGIC-53 transcripts were remarkably altered after V. harveyi challenge, suggesting putative function in host innate immunity.

Supplementary information: The online version contains supplementary material available at 10.1007/s10499-022-00845-3.

Keywords: CTL; ERGIC-53; Immune response; Macrobrachium rosenbergii; V. harveyi.