In Silico Genomics of Fungal Oxidoreductase Genes
Year: 2015
Type: dissertation
Abstract: Oxidoreductases are found in all living organisms and play essential roles in housekeeping, perception of environmental stress, plant-pathogen interactions, defense reactions, and pathogenicity. In particular, laccase, peroxidases and NADPH oxidase, have been implicated in virulence of phytopathogenic fungi in pathogenicity. Despite its relevance towards plant microbe interaction, the identification and comparative analysis of fungal peroxidase-encoding genes at the genomic level have been limited by the lack of a bioinformatics platform as well as paucity of information on transcript profiling of potential candidate oxidoreductase genes. In this project, one of the primary tasks addressed was the construction and development of a new fungal peroxidase database (fPoxDB). The availability of fPoxDB platform facilitated comparative and evolutionary studies of fungal peroxidases at the genomic level. The database contains 6,113 peroxidase genes of 25 gene families from 331 genomes. The archived genes were subjected to pre-computational analyses using eight different bioinformatic tools: SignalP 3.0, SecretomeP 1.0f, TargetP 1.1b, predictNLS, ChloroP 1.1, TMHMM 2.0c, PSortII, and InterPro Scan. Similarity search tools including HMMER, ClustalW, Blast, and BlastMatrix are provided on the platform. A web interface enables researchers to browse the database via either species or class . Graphics support the work and enhance user-friendliness. Retrieved data can be exported to other family web-systems including the comparative fungal genomics platform (CFGP). In parallel to the above study, the second project was focused on in silico comparative analysis fungal NADPH oxidase (Nox) genes sequences. Nox proteins are transmembrane enzymes found in most eukaryotic organisms and influence many biological processes by generating reactive oxygen species (ROS). In fungi, Nox enzymes play roles in pathogenicity, such as the weakening of plant cell walls by ROS. The enzymes exhibit high sequence similarities to the ferric-reductases (Fre) and ferric-chelate reductases (FRO) proteins, which are involved in reduction of Fe3+ to Fe2+ for iron uptake. A total of 34 eukaryotic genomes, covering 28 fungal, one Oomycota, three animal and two plant species, were subjected to bioinformatic analysis. The results indicate that the properties of fungal Nox genes differ from those of the human and plants, providing novel insights that will enable more accurate identification and characterization of the fungal genes. In the third project, as not much is known is about precise role of oxidoreductase encoding genes in pathogenesis, we explored expression profiling of a typical oxidoreductase gene, laccase, in Heterobasidion conifer pathosystem. Laccases are multi-copper oxidoreductases catalyzing the oxidation of phenolic substrates, and they play diverse roles in plants and fungi. In fungi, laccase have been shown to be involved in pathogenicity, as well as in lignin degradation. To understand the potential roles of laccases of the forest pathogen Heterobasidion annosum, a total of 18 laccase genes was identified in this fungus and phylogenetically analyzed. The expression levels of the genes, and laccase activities, during growth of H. annosum on its host in the presence or absence of additional carbon source, such as glucose and sucrose, were investigated. Based on increased transcript expression levels eight laccases were considered to be potentially involved in H. annosum virulence. In summary, we provide the research community with a database dedicated to fungal peroxidases, and our in silico analysis affords new insights in the structure of fungal NADPH oxidases. Lastly, we present experimental evidence that some H. annosum laccases might be involved in virulence during infection of non-suberized Scots pine seedlings.
Source: Työväentutkimus Vuosikirja
Author: Nicolas Détry
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W1699263402