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Fusarium moniliforme secretes four endopolygalacturonases derived from a single gene product

Caprari, Claudio and Bergmann, C. and Migheli, Quirico and Salvi, Giovanni and Albersheim, Peter and Darvill, Alan and Cervone, Felice and De Lorenzo, Giulia (1993) Fusarium moniliformesecretes four endopolygalacturonases derived from a single gene product. Physiological and Molecular Plant Pathology, Vol. 43 (6), p. 453-462. ISSN 0885-5765. Article.

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DOI: 10.1006/pmpp.1993.1073

Abstract

Extracellular endopolygalacturonase, purified from the pathogenic fungus Fusarium moniliforme, consists of four molecular forms (38, 41·5, 45, and 48·5 kDa, respectively. Three forms (38, 41·5, and 45 kDa) were purified to homogencity by FPLC on a Mono S column followed by electroelution after SDS-PAGE. The N-terminal amino acid sequences of each of the three forms, and of a mixture containing all four forms were shown to be identical to that predicted from the nucleotide sequence of the endopolygalacturonase gene previously cloned from F. moniliforme. Enzymatic deglycosylation experiments revealed the presence of N-linked, high mannose oligosaccharide side-chains on all four forms of endo polygalacturonase. Hydrogen fluoride catalysed chemical deglycosylation of the polygalacturonase mixture yielded a single polypeptide with an apparent molecular mass of 36·2 kDa. Southern blot analysis, carried out at high stringency with an endopolygalacturonase-specific probe on genomic DNA digested with three different restriction enzymes, showed a single hybridizing restriction fragment in all three digests. A single 2·0 Mb chromosome hybridized with the endo polygalacturonase-specific probe, as shown by Southern blot analysis of F. moniliforme chromosomes separated by CHEF electrophoresis. Northern blot analysis revealed only one mRNA species 1350 nt encoding endo polygalacturonase. These data indicate that a single gene encodes the endopolygalacturonases of F. moniliforme.

Item Type:Article
ID Code:260
Status:Published
Refereed:Yes
Uncontrolled Keywords:FPLC, fast protein liquid chromatography, CHEF, contour-clamped homogeneous electric field
Subjects:Area 07 - Scienze agrarie e veterinarie > AGR/12 Patologia vegetale
Divisions:001 Università di Sassari > 01 Dipartimenti > Protezione delle piante
Publisher:Academic Press / Elsevier
ISSN:0885-5765
Copyright Holders:© 1993 Academic Press
Deposited On:18 Aug 2009 10:01

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