Molecular characterization of a subtilase from the vascular wilt fungus Fusarium oxysporum

Molecular Plant-microbe Interactions : MPMI
A Di PietroM Isabel G Roncero

Abstract

The gene prt1 was isolated from the tomato vascular wilt fungus Fusarium oxysporum f. sp. lycopersici, whose predicted amino acid sequence shows significant homology with subtilisin-like fungal proteinases. Prt1 is a single-copy gene, and its structure is highly conserved among different formae speciales of F. oxysporum. Prt1 is expressed constitutively at low levels during growth on different carbon and nitrogen sources and strongly induced in medium containing collagen and glucose. As shown by reverse transcription-polymerase chain reaction and fluorescence microscopy of F. oxysporum strains carrying a prt1-promoter-green fluorescent protein fusion, prt1 is expressed at low levels during the entire cycle of infection on tomato plants. F. oxysporum strains transformed with an expression vector containing the prt1 coding region fused to the inducible endopolygalacturonase pg1 gene promoter and grown under promoter-inducing conditions secreted high levels of extracellular subtilase activity that resolved into a single peak of pI 4.0 upon isoelectric focusing. The active fraction produced two clearing bands of 29 and 32 kDa in sodium dodecyl sulfate gels containing gelatin. Targeted inactivation of prt1 in F. oxysporum f. sp. lyc...Continue Reading

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Citations

Jan 30, 2003·Fungal Genetics and Biology : FG & B·Laurence V BindschedlerRichard M Cooper
Feb 16, 2002·European Journal of Biochemistry·Anja I PekkarinenMarja-Leena Niku-Paavola
Jul 16, 2005·Food Additives and Contaminants·A SantinoG Mita
May 15, 2007·Current Opinion in Infectious Diseases·Rod J Hay
Sep 1, 2003·Molecular Plant Pathology·Antonio Di PietroM Isabel G Roncero
Apr 30, 2009·Molecular Plant Pathology·Caroline B Michielse, Martijn Rep
Mar 1, 2007·Molecular Plant Pathology·Petra M HoutermanMartijn Rep
Mar 4, 2015·Proceedings of the National Academy of Sciences of the United States of America·Braham DhillonRichard C Hamelin
Apr 16, 2003·Molecular Microbiology·Zaira CaracuelAntonio Di Pietro
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Jan 4, 2005·Biochemistry. Biokhimii︠a︡·T A Valueva, V V Mosolov
Jul 1, 2003·The New Phytologist·Ghislaine RecorbetClaude Alabouvette

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