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Cloning and expression of selenocysteine methyltransferase cDNA from Camellia sinensis

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Abstract

Selenocysteine methyltransferase (SMT), specifically methylates selenocysteine (SeCys) to produce the nonprotein amino acid Se-methyl selenocysteine (SeMSC) and played key role of removing selenium toxic effect at higher levels to the plant. Here we report the cloning of a cDNA encoding selenocysteine methyltransferase from Camellia sinensis (CsSMT) and expression of CsSMT in Escherichia coli. CsSMT isolated by RT-PCR and RACE-PCR reaction. CsSMT is a 1,401 bp cDNA with an open reading frame predicted to encode a 351 amino acid, 40.5 kDa protein; The predicted amino acid sequences of CsSMT shows 74% identity with A. bisulcatus selenocysteine methyltransferase (AbSMT) and 69% identity with Broccoli (Brassica oleracea var. italica) selenocysteine methyltransferase (BoSMT), and shares 53, 73 and 65% identity, respectively, with Arabidopsis thaliana homocysteine S-methyltransferase AtHMT1, AtHMT2, and AtHMT3, and 65% to Zea mays homocysteine S-methyltransferase (ZmHMT2). Analyses of CsSMT showed that it lacks obvious chloroplast or mitochondrial targeting sequences and contains a consensus sequence of GGCC for a possible zinc-binding motif near the C-terminal and a conserved Cys residue upstream of the zinc-binding motif as other related methyltransferases. Expression of CsSMT correlated with the presence of SMT enzyme activity in cell extracts, and bacteria containing recombinant CsSMT plasmid showed much high tolerance to selenate and selenite.

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Abbreviations

AbSMT:

A. bisulcatus selenocysteine methyltransferase

AtHM:

Arabidopsis thaliana homocysteine S-methyltransferase

BoSMT:

Brassica oleracea var. italica selenocysteine methyltransferase

bp:

Base pair

CsSMT:

Camellia sinensis selenocysteine methyltransferase

HMT:

Homocysteine S-methyltransferase

kb:

Kilobase pair(s)

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcription polymerase chain reaction

SeCys:

Selenocysteine

SeMSC:

Se-methyl selenocysteine

SMT:

Selenocysteine methyltransferase

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Acknowledgments

The authors thank the Director, Key Laboratory of Tea Biochemistry and Biotechnology, Zheng-Zhu Zhang for providing facilities. We thankfully acknowledges the financial support provided by Anhui Provincial Natural Science Foundation, China (grant No.050410102)

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Authors and Affiliations

  1. Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University China, Changjiang West Road 130, Hefei, 230036, Anhui, China

    Lin Zhu, Chang-Jun Jiang, Wei-Wei Deng, Xuan Gao, Rang-Jian Wang & Xiao-Chun Wan

  2. College of Resources and Environmental Sciences, Anhui Agricultural University China, Changjiang West Road 130, Hefei, 230036, Anhui, China

    Lin Zhu

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  1. Lin Zhu
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  2. Chang-Jun Jiang
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  3. Wei-Wei Deng
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  4. Xuan Gao
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Correspondence to Chang-Jun Jiang.

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Communicated by J. Sadowski.

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Zhu, L., Jiang, CJ., Deng, WW. et al. Cloning and expression of selenocysteine methyltransferase cDNA from Camellia sinensis . Acta Physiol Plant 30, 167–174 (2008). https://doi.org/10.1007/s11738-007-0105-7

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  • DOI: https://doi.org/10.1007/s11738-007-0105-7

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