title: An anti-apoptotic B-cell lymphoma-2 (BCL-2) from Channa striatus: Sequence analysis and delayed and advanced gene expression in response to fungal, bacterial and poly I:C induction.
abstract: B-cell lymphoma-2 (BCL-2) is a suppressor of apoptosis and inhibits the caspase dependent apoptosis pathway. In this study, we report molecular characterization of a cDNA sequence encoded of BCL-2 from striped murrel, Channa striatus. A partial cDNA sequence of CsBCL-2 was identified from the striped murrel cDNA library during annotation. Subsequently, the full length CsBCL-2 cDNA sequence was obtained by an internal sequencing method using a forward primer. The sequence contains 699 nucleotide base pairs which encode 232 amino acid residues. The domain and motif analysis revealed that the CsBCL-2 polypeptide consists of BCL-2 homologous domain BH4 at the N-terminal region between 4 and 21 and the BCL-2 homologous domains BH1, BH2 and BH3 between 87 and 187. The CsBCL-2 polypeptide sequence does not have a signal peptide region, but it consists of two novel transmembrane regions at 134-152 and 209-226. The sequence analysis showed that the CsBCL-2 has highest sequence identity (70%) with BCL-2 like protein 1 (BCL-2 L1) from pufferfish Takifugu rubripes. The phylogenetic analysis showed that the CsBCL-2 was situated in the BCL-2 L1 fish clade. The secondary analysis showed that the CsBCL-2 protein consists of 132 amino acid residues in the α-helical region and 100 amino acid residues in the random coil region. The validated 3D structure of CsBCL-2 showed the active residues Gly(135) and Arg(136) in the 7th α-helical position, whereas Trp(178) is in the 9th α-helical region. CsBCL-2 mRNA transcription is predominately present in spleen and is upregulated upon being induced with fungus Aphanomyces invadans, bacteria Aeromonas hydrophila, Escherichia coli LPS, Laminaria digitata beta-1,3-glucan and poly I:C. Overall, the CsBCL-2 mRNA transcription results indicate the potential involvement of CsBCL-2 in immune system of C. striatus. However, further research at proteomic level is necessary to examine these predictions.
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India. Electronic address: jesuaraj@hotmail.com.
authors.forename: Jesu
authors.initials: J
authors.lastname: Arockiaraj
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Rajesh
authors.initials: R
authors.lastname: Palanisamy
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India; Department of Microbiology, SRM Arts & Science College, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Abirami
authors.initials: A
authors.lastname: Arasu
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India; Department of Biotechnology, SRM Arts & Science College, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Akila
authors.initials: A
authors.lastname: Sathyamoorthi
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Venkatesh
authors.initials: V
authors.lastname: Kumaresan
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Prasanth
authors.initials: P
authors.lastname: Bhatt
authors.affiliation: Division of Fisheries Biotechnology & Molecular Biology, Research Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai 603203, Tamil Nadu, India.
authors.forename: Mukesh Kumar
authors.initials: MK
authors.lastname: Chaurasia
authors.affiliation: Lab PCN 206, Microbiology Division, CSIR-Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, Uttar Pradesh, India.
authors.forename: Mukesh
authors.initials: M
authors.lastname: Pasupuleti
authors.affiliation: Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, TX 78712, USA.
authors.forename: Annie J
authors.initials: AJ
authors.lastname: Gnanam
mesh_terms: D016980:Aeromonas hydrophila / Q000187:drug effects / Q000502:physiology; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D044744:Aphanomyces / Q000187:drug effects; D017209:Apoptosis / Q000235:genetics*; D001483:Base Sequence; D019295:Computational Biology; D005786:Gene Expression Regulation / Q000187:drug effects; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics* / Q000382:microbiology*; D010802:Phylogeny; D011070:Poly I-C / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D020539:Sequence Analysis, Protein
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D011070:Poly I-C
keywords: B-cell lymphoma; Bioinformatics; Channa striatus; Delayed response; Gene expression