Expression cloning of a fungal proline-rich glycoprotein specific to the biotrophic interface formed in the Colletotrichum-bean interaction
| Autor: | Sarah E. Perfect, Elaine F. Green, Christiane Doering-Saad, Richard J. O'Connell, Jonathan R. Green |
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| Rok vydání: | 1998 |
| Předmět: |
Signal peptide
DNA Complementary DNA Plant Proline Transcription Genetic Molecular Sequence Data Plant Science Transfection Fungal Proteins Cell wall Ascomycota Complementary DNA Genetics Animals Amino Acid Sequence Peptide sequence Glycoproteins chemistry.chemical_classification Plants Medicinal Base Sequence biology Colletotrichum lindemuthianum fungi Antibodies Monoclonal Fabaceae Cell Biology biology.organism_classification Recombinant Proteins Biochemistry Colletotrichum chemistry Protein Biosynthesis COS Cells Expression cloning Glycoprotein |
| Zdroj: | The Plant Journal. 15:273-279 |
| ISSN: | 1365-313X 0960-7412 |
| DOI: | 10.1046/j.1365-313x.1998.00196.x |
| Popis: | The monoclonal antibody, UB25, recognises a glycoprotein specifically located at the biotrophic interface formed in the Colletotrichum lindemuthianum-bean interaction. The antibody labels the walls of intracellular hyphae and the interfacial matrix which separates them from the invaginated host plasma membrane. In Western blots, UB25 recognises a ladder of bands which are multiples of M(r) 40.5 kDa. A full length cDNA encoding the glycoprotein recognised by UB25 has been isolated by expression cloning and designated CIH1 (Colletotrichum Intracellular Hypha 1). In vitro transcription/translation of CIH1, and transfection of mammalian COS cells, showed that UB25 recognized the expressed product in both procedures confirming that the clones isolated were true positives. Southern analysis of bean and C. lindemuthianum genomic DNA indicated that the CIH1 glycoprotein is fungally encoded and Northern analysis showed that it is only expressed in planta. Analysis of the deduced amino acid sequence of CIH1 indicates the presence of an N-terminal signal sequence and two possible sites for N-glycosylation. The N-terminal domain of the mature protein is rich in proline and contains several short repetitive motifs. CIH1 is thus a fungal proline-rich glycoprotein which appears to form a cross-linked structure in planta and, as such, resembles plant cell wall proline- and hydroxyproline-rich proteins. Possible functions for the CIH1 protein in the establishment and maintenance of biotrophy are discussed. |
| Databáze: | OpenAIRE |
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