clap1, a gene encoding a copper-transporting ATPase involved in the process of infection by the phytopathogenic fungus Colletotrichum lindemuthianum

Abstract

A screen for insertional mutants of Colletrichum lindemuthianum, the causative agent of common bean anthracnose, led to the identification of a non-pathogenic, lightly colored transformant. This mutant is unable to induce disease symptoms on intact or wounded primary leaves of seedlings and plantlets of Phaseolus vulgaris. In vitro, it exhibits normal vegetative growth, sporulation and conidial germination, but the cultures remain beige instead of becoming black. Microscopic examination revealed that this mutant forms fewer appressoria than the wild-type strain, and these are misshapen and poorly melanized. Molecular analyses indicated that the mutagenic plasmid had targeted clap1, a gene encoding a putative copper-transporting ATPase sharing 35% identity with the human Menkes and Wilson proteins and the product of the CCC2 gene of Saccharomyces cerevisiae. Complementation of the non-pathogenic beige mutant with a wild-type allele of clap1 restored both pathogenicity and pigmentation. Conversely, replacement of the wild-type allele with a disrupted clap1 gene gave rise to non-pathogenic beige transformants. Compared with the wild-type strain, extracts from clap1 mutants were found to have very low levels of phenol oxidase activity. These observations suggest that the clap1 gene product may be involved in the pathogenicity of C. lindemuthianum strains because of its role in delivering copper to secreted cuproenzymes, such as the phenol oxidases that mediate the polymerization of 1,8-dihydroxynaphthalene to melanin.

Identification of soluble secreted proteins from appressoria of Colletotrichum higginsianum by analysis of expressed sequence tags

“…In the present study, we identified two putative secreted laccases that differed in amino acid sequence (data not shown), suggesting that C. higginsianum deploys at least two laccase isoenzymes. We also found a homologue of the copper-transporting ATPase CLAP1, which is required for appressorial melanization in C. lindemuthianum and is thought to deliver copper to secreted cuproenzymes such as laccases (Parisot et al, 2002).…”

Contribution of ATPase copper transporters in animal but not plant virulence of the crossover pathogen Aspergillus flavus

“…Although many studies have shown that the Cu detoxification system plays important roles in virulence of human pathogens [15,48,49], to date there hasn’t been a study of a role for CrpA in a plant pathogenic fungus, nor is there any evidence the plant cells use Cu as a defense mechanism. The only study investigating any role of a Cu transporter in a plant pathogen is where a Cu golgi transporter, BcCCC2, in the pathogen Botrytis cinerea was found important for virulence in that fungus [24].…”

Identification of a Copper-Inducible Promoter for Use in Ectopic Expression in the Fungal Pathogen Histoplasma capsulatum

“…In the fungal pathogen C. neoformans, activity of the melanin biosynthetic enzyme laccase, which is required for virulence (30), is influenced by copper levels (42). Other data imply a more direct role for copper transporters during infection of the host; for example, clap1, the ortholog of H. capsulatum CRP1 in the phytopathogenic fungus C. lindemuthianum, is required for pathogenesis (25). Similarly, mutation of CtpA, a coppertransporting P-type ATPase in Listeria monocytogenes, results in a marked reduction in host colonization (9).…”