Document Detail

Alternative splicing: a novel mechanism of regulation identified in the chorismate mutase gene of the potato cyst nematode Globodera rostochiensis.
MedLine Citation:
PMID:  18786575     Owner:  NLM     Status:  MEDLINE    
Chorismate mutase (CM) secreted from the stylet of plant-parasitic nematodes plays an important role in plant parasitism. We isolated and characterized a new nematode CM gene (Gr-cm-1) from the potato cyst nematode, Globodera rostochiensis. The Gr-cm-1 gene was found to exist in the nematode genome as a single-copy gene that has two different alleles, Gr-cm-1A and Gr-cm-1B, both of which could give rise to two different mRNA transcripts of Gr-cm-1 and Gr-cm-1-IRII. In situ mRNA hybridization showed that the Gr-cm-1 gene was exclusively expressed within the subventral oesophageal gland cells of the nematode. Gr-cm-1 was demonstrated to encode a functional CM (GR-CM-1) potentially having a dimeric structure as the secreted bacterial *AroQ CMs. Gr-cm-1-IRII, generated by retention of intron 2 of the Gr-cm-1 pre-mRNA through alternative splicing (AS), would encode a truncated protein (GR-CM-1t) lacking the CM domain with no CM activity. The quantitative real-time reverse transcription-PCR assay revealed that splicing of the Gr-cm-1 gene was developmentally regulated; Gr-cm-1 was up-regulated whereas Gr-cm-1-IRII was down-regulated in early nematode parasitic stages compared to the preparasitic juvenile stage. Low-temperature SDS-PAGE analysis revealed that GR-CM-1 could form homodimers when expressed in Escherichia coli and the dimerization domain was retained in the truncated GR-CM-1t protein. The specific interaction between the two proteins was demonstrated in yeast. Our data suggested that the novel splice variant might function as a dominant negative isoform through heterodimerization with the full-length GR-CM-1 protein and that AS may represent an important mechanism for regulating CM activity during nematode parasitism.
Shun-Wen Lu; Duanhua Tian; Harmony B Borchardt-Wier; Xiaohong Wang
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Publication Detail:
Type:  Journal Article     Date:  2008-08-22
Journal Detail:
Title:  Molecular and biochemical parasitology     Volume:  162     ISSN:  0166-6851     ISO Abbreviation:  Mol. Biochem. Parasitol.     Publication Date:  2008 Nov 
Date Detail:
Created Date:  2008-09-23     Completed Date:  2008-12-10     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8006324     Medline TA:  Mol Biochem Parasitol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1-15     Citation Subset:  IM    
Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, NY 14853, USA.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/EF437153;  EF437154;  EF437155;  EF437156
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MeSH Terms
Alternative Splicing*
Amino Acid Sequence
Base Sequence
Chorismate Mutase / genetics,  metabolism*
Escherichia coli / enzymology,  genetics
Gene Expression Regulation, Developmental*
Host-Parasite Interactions
Introns / genetics
Molecular Sequence Data
Plant Diseases / parasitology
Protein Isoforms
Sequence Analysis, DNA
Solanum tuberosum / parasitology*
Two-Hybrid System Techniques
Tylenchoidea / enzymology*,  genetics,  metabolism,  physiology*
Reg. No./Substance:
0/Protein Isoforms; EC Mutase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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