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Characterization of a plant (rice) translin and its comparative analysis with human translin

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Abstract

Main conclusion

For the first time, a plant (rice) translin was characterized. The rice translin protein, which was octameric in native state, bound efficiently to single-stranded DNA and RNA.

Translin, a DNA-/RNA-binding protein, is expressed in brain, testis and in certain malignancies. It is involved in chromosomal translocation, mRNA metabolism, transcriptional regulation and telomere protection. Studies from human, mice, drosophila and yeast have revealed that it forms an octameric ring, which is important for its function. In spite of the absence of neuronal functions and cancer processes, translin is present in plant systems, but information on plant translin is lacking. Here we report the characterization of a plant (rice) translin. Translin cDNA from O. sativa was cloned into an expression vector; protein was over-expressed in E. coli and subsequently purified to homogeneity. Circular dichroism and homology-based modeling showed that the rice translin protein was similar to the other translin proteins. Native PAGE and gel-filtration analyses showed rice translin to form an octamer and this octameric assembly was independent of disulphide bonds. Rice translin bound to single-stranded DNA sequences like human translin, but not to the double-stranded DNA. Rice translin bound more efficiently to linear DNA (with staggered ends) than open or closed circular DNA. Rice translin also bound to RNA, like its human counterpart. Rice translin displays all the characteristic properties of the translin group of proteins and does indeed qualify as a bonafide “translin” protein. To our knowledge, this is the first report wherein the translin protein from a plant source has been functionally characterized. Understanding the translin biology from plant systems will give the new insights into its functional role during plant development.

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Abbreviations

HsTsn:

Homo sapiens translin

OsTsn:

Oryza sativa translin

TB-RBP:

Testis, brain RNA-binding protein

TRAX:

Translin-associated factor-X

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Acknowledgments

We sincerely acknowledge Rice Genome Resource Center (RGRC), Japan for providing translin cDNA clone. Our thanks are due to Dr. Anubrata Das, for help with the homology-based modeling and Dr. Himanshi Narang Mishra, for the help during cloning of translin cDNA. We acknowledge Mr. Shivam Shukla for his help in gel-filtration experiment. Dr. S. K.Apte, Head, Molecular Biology Division, Bhabha Atomic Research Center, Mumbai, India, is also acknowledged for his constant support throughout the study. We acknowledge Dr. Vinay Kumar, Protein Crystallography Section, Solid State Physics Division, Bhabha Atomic Research Center, Mumbai, India, for helpful suggestions and critical reviewing of the manuscript. The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Biomolecular Damage and Repair Section, Molecular Biology Division, Bhabha Atomic Research Center, Trombay, Mumbai, 400085, India

    Rajani Kant Chittela & Anand Ballal

  2. Protein Crystallography Section, Solid State Physics Division, Bhabha Atomic Research Center, Trombay, Mumbai, 400085, India

    Gagan Deep Gupta

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  1. Rajani Kant Chittela
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Correspondence to Rajani Kant Chittela.

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Chittela, R.K., Gupta, G.D. & Ballal, A. Characterization of a plant (rice) translin and its comparative analysis with human translin. Planta 240, 357–368 (2014). https://doi.org/10.1007/s00425-014-2092-4

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