Identification of two suites of cyclotide precursor genes from metallophyte Viola baoshanensis: cDNA sequence variation, alternative RNA splicing and potential cyclotide diversity

Zhang, J, Liao, B, Craik, D. J., Li, J. T., Hu, M and Shu, W. S. (2009) Identification of two suites of cyclotide precursor genes from metallophyte Viola baoshanensis: cDNA sequence variation, alternative RNA splicing and potential cyclotide diversity. GENE, 431 1-2: 23-32. doi:10.1016/j.gene.2008.11.005


Author Zhang, J
Liao, B
Craik, D. J.
Li, J. T.
Hu, M
Shu, W. S.
Title Identification of two suites of cyclotide precursor genes from metallophyte Viola baoshanensis: cDNA sequence variation, alternative RNA splicing and potential cyclotide diversity
Journal name GENE   Check publisher's open access policy
ISSN 0378-1119
Publication date 2009-02-15
Year available 2008
Sub-type Article (original research)
DOI 10.1016/j.gene.2008.11.005
Volume 431
Issue 1-2
Start page 23
End page 32
Total pages 10
Editor G Bernardi
Place of publication Amsterdam. Netherlands
Publisher Elsevier BV
Language eng
Subject C1
970106 Expanding Knowledge in the Biological Sciences
060702 Plant Cell and Molecular Biology
Abstract Cyclotides are a novel family of plant-derived defense peptides that are biosynthetically produced via the processing of cyclotide precursor (CP) proteins containing one, two or three cyclotide domains. By screening a cDNA library of Viola baoshanensis roots and using RACE and RT-PCR methods, 23 cDNA clones were identified and then used to deduce full CP proteins containing one (VbCP1S-5), two (VbCP6S), or three (VbCP7S) cyclotide domains. RT-PCR and sequence analyses suggested that VbCP6S were resulted from the alternative splicing of VbCP7S RNA. The significance of VbCP7S RNA splicing is that it provides a mechanism for increasing the diversity of cyclotide expression via the recombination of N-terminal repeat (NTR) regions and cyclotide domains. After analyzing the full endoplasmic reticulum (ER) signals of known and novel CPs associated with RT-PCR tests, three primers encoding the conserved sequence ALVLIATFA, AAFALPA-LA and AAFALPA-AFA were proposed to be more efficient in cloning CP genes than the well-applied primer encoding AAFALPA. Cyclotide sequence analyses indicated that the cDNA clones encoded a variety of Möbius and bracelet cyclotides, which were likely involved in the known bioactivities of cyclotides, and also might play a previously unreported role in mediating the metal tolerance of V. baoshanensis. Overall, this study shows that CP genes are varied in V. baoshanensis and cyclotide expression is subject to transcriptional and post-transcriptional regulation in this plant.
Keyword Alternative RNA splicing
Cyclic Cystine Knot
Cyclotide
Cyclotide diversity
Cyclotide precursor
Sequence Variation
Viola baoshanensis
Q-Index Code C1
Q-Index Status Provisional Code
Additional Notes Available online 19 November 2008

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
 
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Created: Thu, 03 Sep 2009, 08:45:54 EST by Mr Andrew Martlew on behalf of Institute for Molecular Bioscience