Immunological role of C4 CC chemokine-1 from snakehead murrel Channa striatus.
In this study, we have reported a cDNA sequence of C4 CC chemokine identified from snakehead murrel (also known as striped murrel) Channa striatus (named as CsCC-Chem-1) normalized cDNA library constructed by Genome Sequencing FLX™ Technology (GS-FLX™). CsCC-Chem-1 is 641 base pairs (bp) long that contain 438 bp open reading frame (ORF). The ORF encodes a polypeptide of 146 amino acids with a molecular mass of 15 kDa. The polypeptide contains a small cytokine domain at 30-88. The domain carries the CC motif at Cys(33)-Cys(34). In addition, CsCC-Chem-1 consists of another two cysteine residues at C(59) and C(73), which, together with C(33) and C(34), make CsCC-Chem-1 as a C4-CC chemokine. CsCC-Chem-1 also contains a 'TCCT' motif at 32-35 as CC signature motif; this new motif may represent new characteristic features, which may lead to some unknown function that needs to be further focused on. Phylogenitically, CsCC-Chem-1 clustered together with CC-Chem-1 from rock bream Oplegnathus fasciatus and European sea bass Dicentrarchus labrax. Significantly (P<0.05) highest gene expression was noticed in spleen and is up-regulated upon fungus (Aphanomyces invadans), bacteria (Aeromonas hydrophila) and virus (poly I:C) infection at various time points. The gene expression results indicate the influence of CsCC-Chem-1 in the immune system of murrel. Overall, the gene expression study showed that the CsCC-Chem-1 is a capable gene to increase the cellular response against various microbial infections. Further, we cloned the coding sequence of CsCC-Chem-1 in pMAL vector and purified the recombinant protein to study the functional properties. The cell proliferation activity of recombinant CsCC-Chem-1 protein showed a significant metabolic activity in a concentration dependent manner. Moreover, the chemotaxis assay showed the capability of recombinant CsCC-Chem-1 protein which can induce the migration of spleen leukocytes in C. striatus. However, this remains to be verified further at molecular and proteomic level.
Division of Fisheries Biotechnology & Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603 203, Tamil Nadu, India.
Prasanth
P
Bhatt
Venkatesh
V
Kumaresan
Rajesh
R
Palanisamy
Mukesh Kumar
MK
Chaurasia
Annie J
AJ
Gnanam
Mukesh
M
Pasupuleti
Jesu
J
Arockiaraj
D016980:Aeromonas hydrophila / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D044744:Aphanomyces / Q000276:immunology*; D001483:Base Sequence; D002465:Cell Movement; D049109:Cell Proliferation; D019742:Chemokines, CC / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D007962:Leukocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D010473:Perciformes / Q000276:immunology*; D010802:Phylogeny; D011070:Poly I-C / Q000276:immunology*; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000096:biosynthesis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D019742:Chemokines, CC; D029941:Fish Proteins; D012333:RNA, Messenger; D011070:Poly I-C
Cell proliferation; Chemokine; Chemotaxis; Epizootic ulcerative syndrome (EUS); Murrel