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Plant Cell Advance Online Publication
Published on August 25, 2006; 10.1105/tpc.106.042812

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Received April 11, 2006
Returned for revision June 1, 2006
Accepted July 26, 2006

Discovery of Cyclotide-Like Protein Sequences in Graminaceous Crop Plants: Ancestral Precursors of Circular Proteins?

Jason P. Mulvenna 1, Joshua S. Mylne 1, Rekha Bharathi 1, Rachel A. Burton 2, Neil J. Shirley 2, Geoffrey B. Fincher 2, Marilyn A. Anderson 3, and David J. Craik 1*

1 Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland 4072, Australia
2 Australian Centre for Plant Functional Genomics, School of Agriculture and Wine, University of Adelaide, Waite Campus, Glen Osmond, South Australia 5064, Australia
3 Department of Biochemistry, La Trobe University, Melbourne, Victoria 3086, Australia

* To whom correspondence should be addressed. E-mail: d.craik{at}imb.uq.edu.au.

Cyclotides are peptides from plants of the Rubiaceae and Violaceae families that have the unusual characteristic of a macrocylic backbone. They are further characterized by their incorporation of a cystine knot in which two disulfides, along with the intervening backbone residues, form a ring through which a third disulfide is threaded. The cyclotides have been found in every Violaceae species screened to date but are apparently present in only a few Rubiaceae species. The selective distribution reported so far raises questions about the evolution of the cyclotides within the plant kingdom. In this study, we use a combined bioinformatics and expression analysis approach to elucidate the evolution and distribution of the cyclotides in the plant kingdom and report the discovery of related sequences widespread in the Poaceae family, including crop plants such as rice (Ozyza sativa), maize (Zea mays), and wheat (Triticum aestivum), which carry considerable economic and social importance. The presence of cyclotide-like sequences within these plants suggests that the cyclotides may be derived from an ancestral gene of great antiquity. Quantitative RT-PCR was used to show that two of the discovered cyclotide-like genes from rice and barley (Hordeum vulgare) have tissue-specific expression patterns.




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