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

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Abstract

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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Abbreviations

CsmMnSOD:

Channa striatus mitochondrial manganese superoxide dismutase

ROS:

Reactive oxygen species

–OH:

Hydroxyl radical

H2O2 :

Hydrogen peroxide

O2 − :

Superoxide radical

Cu/ZnSOD:

Copper/zinc SOD

FeSOD:

Iron SOD

NiSOD:

Nickel SOD

MTS:

Mitochondrial translocation sequence

EUS:

Epizootic ulcerative syndrome

MTP:

Mitochondrial targeting peptide

FPS:

Fish physiological saline

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Acknowledgments

This research is supported by DBT’s Prestigious Ramalingaswami Re-entry Fellowship (D.O.NO.BT/HRD/35/02/2006) funded by Department of Biotechnology, Ministry of Science and Technology, Government of India, New Delhi.

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Authors and Affiliations

  1. Division of Fisheries Biotechnology and Molecular Biology, Department of Biotechnology, Faculty of Science and Humanities, SRM University, Kattankulathur, Chennai, 603 203, Tamil Nadu, India

    Jesu Arockiaraj, Rajesh Palanisamy, Prasanth Bhatt & Venkatesh Kumaresan

  2. Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, TX, 78712, USA

    Annie J. Gnanam

  3. 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

    Mukesh Pasupuleti

  4. Department of Biotechnology, Faculty of Applied Sciences, AIMST University, Semeling Bedong, 08100, Bedong, Kedah, Malaysia

    Marimuthu Kasi

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  1. Jesu Arockiaraj
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  2. Rajesh Palanisamy
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Arockiaraj, J., Palanisamy, R., Bhatt, P. et al. A novel murrel Channa striatus mitochondrial manganese superoxide dismutase: gene silencing, SOD activity, superoxide anion production and expression. Fish Physiol Biochem 40, 1937–1955 (2014). https://doi.org/10.1007/s10695-014-9981-0

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