An upstream initiator caspase 10 of snakehead murrel Channa striatus, containing DED, p20 and p10 subunits: Molecular cloning, gene expression and proteolytic activity

Arockiaraj, Jesu; Gnanam, Annie J.; Dhanaraj, M.; Pasupuleti, Mukesh; Milton, James; Singh, Arun

doi:10.1016/j.fsi.2012.11.040

Cited by 38 publications

(20 citation statements)

References 56 publications

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“…Using 2.5 lg of RNA, the cDNA was synthesized using a SuperScript Ò VILO TM cDNA Synthesis Kit (Life technologies) as suggested by the manufacturer with slight modifications (Abirami et al 2013;Arockiaraj et al 2013a). The isolated cDNA was stored at -20°C for further analysis.…”

Section: Rna Extraction and Cdna Synthesismentioning

confidence: 99%

A novel murrel Channa striatus mitochondrial manganese superoxide dismutase: gene silencing, SOD activity, superoxide anion production and expression

Arockiaraj

Palanisamy

Bhatt

et al. 2014

Fish Physiol Biochem

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We have reported the molecular characterization including gene silencing, superoxide activity, superoxide anion production, gene expression and molecular characterization of a mitochondrial manganese superoxide dismutase (mMnSOD) from striped murrel Channa striatus (named as CsmMnSOD). The CsmMnSOD polypeptide contains 225 amino acids with a molecular weight of 25 kDa and a theoretical isoelectric point of 8.3. In the N-terminal region, CsmMnSOD carries a mitochondrial targeting sequence and a superoxide dismutases (SOD) Fe domain (28-109), and in C-terminal region, it carries another SOD Fe domain (114-220). The CsmMnSOD protein sequence shared significant similarity with its homolog of MnSOD from rock bream Oplegnathus fasciatus (96%). The phylogenetic analysis showed that the CsmMnSOD fell in the clade of fish mMnSOD group. The monomeric structure of CsmMnSOD possesses 9 α-helices (52.4%), 3 β-sheets (8.8%) and 38.8% random coils. The highest gene expression was noticed in liver, and its expression was inducted with fungal (Aphanomyces invadans) and bacterial (Aeromonas hydrophila) infections. The gene silencing results show that the fish that received dsRNA exhibited significant (P < 0.05) changes in expression when compared to their non-injected and fish physiological saline-injected controls. The SOD activity shows that the activity increases with the spread of infection and decreases once the molecule controls the pathogen. The capacity of superoxide anion production was determined by calculating the granular blood cell count during infection in murrel. It shows that the infection influenced the superoxide radical production which plays a major role in killing the pathogens. Overall, this study indicated the defense potentiality of CsmMnSOD; however, further research is necessary to explore its capability at protein level.

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Section: Rna Extraction and Cdna Synthesismentioning

confidence: 99%

A novel murrel Channa striatus mitochondrial manganese superoxide dismutase: gene silencing, SOD activity, superoxide anion production and expression

Arockiaraj

Palanisamy

Bhatt

et al. 2014

Fish Physiol Biochem

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“…The detailed procedure on C. striatus cDNA library construction was described in our earlier studies (Arockiaraj et al, 2013a;Abirami et al, 2013). From the established cDNA library of C. striatus sequence database, we identified a full length cathepsin B gene, which we designated as CsCath B.…”

Section: Cdna Library Construction Identification and Bioinformaticsmentioning

confidence: 99%

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Arockiaraj¹,

Kumaresan²,

Chaurasia³

et al. 2014

Turk. J. Fish. Aquat. Sci.

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In this study, we have reported a full length cDNA of cathepsin B identified from the constructed cDNA library of snakehead murrel Channa striatus by genome sequence FLX technology. . CsCath B exhibited the maximum similarity (87%) with Cath B from mangrove red snapper, Lutjanus argentimaculatus. Phylogenetically, CsCath B is clustered together with the fish groups belonging to perciformes. A predicted 3D model of CsCath B revealed 11 α-helix and 10 β-strands. CsCath B contains higher percentage (10%) of coils due to the presence of many glycine residues (36 residues). The highest gene expression (P<0.05) was noticed in liver. Further, the expression was induced with fungal (Aphanomyces invadans) and bacterial (Aeromonas hydrophila) infections. The predicted antimicrobial region of CsCath B was synthesized to study its antimicrobial property. The peptide exhibited the antimicrobial activity towards Gram negative and Gram positive bacteria. The overall results indicate that CsCath B is a potential molecule for further studies on murrel defense mechanism.

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“…Using 2.5 µg of RNA, first strand cDNA synthesis was carried out using a SuperScript VILO TM cDNA Synthesis Kit (Life technologies) (Arockiaraj et al 2013). The resulting cDNA solution was stored at -20…”

Section: Rna Isolation and Cdna Conversionmentioning

confidence: 99%

A murrel cysteine protease, cathepsin L: bioinformatics characterization, gene expression and proteolytic activity

et al. 2014

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Cathepsin L, a lysosomal endopeptidase, is a member of the peptidase C1 family (papain-like family) of cysteine proteinases that cleave peptide bonds of lysosomal proteins. In this study, we report a cathepsin L sequence identified from the constructed cDNA library of striped murrel Channa striatus (designated as CsCath L) using genome sequencing FLX TM technology. The full-length CsCath L contains three eukaryotic thiol protease domains at positions 134-145, 278-288 and 299-318. Phylogenetic analysis revealed that the CsCath L was clustered together with other cathepsin L from teleosts. The three-dimensional structure of CsCath L modelled by the I-Tasser program was compared with structures deposited in the Protein Data Bank to find out the structural similarity of CsCath L with experimentally identified structures. The results showed that the CsCath L exhibits maximum structural identity with pro-cathepsin L from human. The RNA fold structure of CsCath L was predicted along with its minimum free energy (-471.93 kcal/mol). The highest CsCath L gene expression was observed in liver, which was also significantly higher (P < 0.05) than that detected in other tissues taken for analysis. In order to investigate the mRNA transcription profile of CsCath L during infection, C. striatus were injected with fungus (Aphanomyces invadans) and bacteria (Aeromonas hydrophila) and its expression was up-regulated in liver at various time points. Similar to gene expression studies, the highest CsCath L enzyme activity was also observed in liver and its activity was up-regulated by fungal and bacterial infections.

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An upstream initiator caspase 10 of snakehead murrel Channa striatus, containing DED, p20 and p10 subunits: Molecular cloning, gene expression and proteolytic activity

Cited by 38 publications

References 56 publications

A novel murrel Channa striatus mitochondrial manganese superoxide dismutase: gene silencing, SOD activity, superoxide anion production and expression

A novel murrel Channa striatus mitochondrial manganese superoxide dismutase: gene silencing, SOD activity, superoxide anion production and expression

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A murrel cysteine protease, cathepsin L: bioinformatics characterization, gene expression and proteolytic activity

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