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Expression cloning of a fungal proline‐rich glycoprotein specific to the biotrophic interface formed in the Colletotrichum –bean interaction
Author(s) -
Perfect Sarah E.,
O’Connell Richard J.,
Green Elaine F.,
DoeringSaad Christiane,
Green Jonathan R.
Publication year - 1998
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1998.00196.x
Subject(s) - biology , glycoprotein , complementary dna , colletotrichum lindemuthianum , proline , glycosylation , peptide sequence , microbiology and biotechnology , signal peptide , biochemistry , amino acid , gene , botany , cultivar
Summary 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 I ntracellular H ypha 1). In vitro transcription/translation of CIH1 , and transfection of mammalian COS cells, showed that UB25 recognised 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.