Reconstructing the diversification of subtilisins in the pathogenic fungus Metarhizium anisopliae
| Autor: | Raymond J. St. Leger, Savita Bagga, Gang Hu, Steven E. Screen |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Nonsynonymous substitution
animal structures Subfamily Molecular Sequence Data Metarhizium anisopliae Evolution Molecular Genetics Amino Acid Sequence Subtilisins DNA Fungal Phylogeny Expressed sequence tag Base Composition biology Phylogenetic tree Models Genetic Sequence Homology Amino Acid fungi Subtilisin Genetic Variation General Medicine Sequence Analysis DNA biology.organism_classification Mutation Metarhizium Mitosporic Fungi Sequence Alignment |
| Zdroj: | Gene. 324 |
| ISSN: | 0378-1119 |
| Popis: | Fungi secrete subtilisin proteinases to acquire nutrients and breach host barriers. Here we sought a global characterization of the diversity of subtilisins in the insect pathogen Metarhizium anisopliae . Expressed sequence tag (EST) analyses showed that a broad host range strain of M. anisopliae sf. anisopliae (strain 2575) expressed 11 subtilisins during growth on insect cuticle, the largest number of subtilisins reported from any fungus. Polymerase chain reaction amplified 10 of their orthologs from a second strain with multiple hosts (strain 820) and seven from the locust specialist M. anisopliae sf. acridum (strain 324). Analyses based on sequence similarities and exon–intron structure grouped M. anisopliae subtilisins into four clusters—a class I (“bacterial”) subtilisin (Pr1C), and three clusters of proteinase K-like class II subtilisins: extracellular subfamily 1 (Pr1A, Pr1B, Pr1G, Pr1I and Pr1K), extracellular subfamily 2 (Pr1D, Pr1E, Pr1F and Pr1J) and an endocellular subtilisin (Pr1H). Phylogenetic analysis of homologous sequences from other genera revealed that this subdivision of proteinase K-like subtilisins into three subfamilies preceded speciation of major fungal lineages. However, diversification has continued during the evolution of Metarhizium subtilisins with evidence of gene duplication events after divergence of M. anisopliae sf. anisopliae and M. anisopliae sf. acridum . Comparing alignments and nonsynonymous/synonymous rates for Pr1 isoenzymes within a lineage and between lineages showed that while overall divergence of subtilisins followed neutral expectations, amino acids involved in catalysis were under strong selective constraint. This suggests that each Pr1 paralog contributes to the pathogens fitness. Furthermore, homology modeling predicted differences between the Pr1's in their secondary substrate specificities, adsorption properties to cuticle and alkaline stability, indicative of functional differences. |
| Databáze: | OpenAIRE |
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