Characterization of the protein processing and secretion pathways in a comprehensive set of expressed sequence tags fromTrichoderma reesei

Abstract

Trichoderma reesei is a filamentous fungus widely used as an efficient protein producer and known to secrete large quantities of biomass degrading enzymes. Much work has been done aimed at improving the secretion efficiency of this fungus. It is generally accepted that the major bottlenecks in secretion are protein folding and ornamentation steps in this pathway. In an attempt to identify genes involved in these steps, the 5P ends of 21 888 cDNA clones were sequenced from which a unique set of over 5000 were also 3P sequenced. Using annotation tools Gene Ontology terms were assigned to 2732 of the sequences. Homologs to the majority of Aspergillus niger's Srg genes as well as a number of homologs to genes involved in protein folding and ornamentation pathways were identified.

Spatially Segregated SNARE Protein Interactions in Living Fungal Cells

“…Understanding of the fungal secretion pathway has significantly increased recently. Based on the analysis of sequenced fungal genomes, it is clear that the secretory machinery of filamentous fungi has similar features to that of other eukaryotes (15,16), and some genes encoding the proteins involved in vesicle trafficking have been cloned and characterized (17)(18)(19)(20)(21)(22).…”

Global Carbon Utilization Profiles of Wild-Type, Mutant, and Transformant Strains of Hypocrea jecorina

“…We are interested in metabolic engineering of the ascomycete Hypocrea jecorina (Trichoderma reesei) to increase the production of cellulase and other extracellular enzymes. The genome sequence of this fungus is now available (http://gsphere.lanl.gov/trire1/trire1 .home.html), as are cDNA sequences from mycelia grown on glucose or under cellulase-inducing conditions (5,7,10,24). However, knowledge of the genome-wide similarity of physiological profiles in various transformed strains and mutants often is needed to meaningfully interpret the gene expression data.…”

Insight into Trichoderma reesei’s genome content, organization and evolution revealed through BAC library characterization

“…Gene Ontology terms were assigned to the assembled dataset using merged annotations from BlastX to the GenBank nr protein database, BlastX to the Gene Ontology manually annotated protein sequence database and Interpro analysis as previously described in Diener et al (2004) (Altschul et al, 1990;Mulder et al, 2003), These annotations were loaded into the Amigo browser to aid analysis (Ashburner et al, 2000;Harris et al, 2004).…”

“…Merging of these annotations produced 532 proteins with at least one GO term annotation. A compilation of major GO categories can be seen in Table 3 along with comparison to the GO annotation of an EST dataset using the same protocol (Diener et al, 2004). The major categories of biological process, cellular component, and molecular function contained 462, 314, and 499 sequences annotated to a child GO term within them.…”

“…These enzymes are frequently used in the food, textile, and paper industries for processing of raw materials (Biely and Tenkanen, 1998). A large number of these enzymes have been previously identified and characterized amongst T. reeseiÕs complement of genes (Diener et al, 2004;Foreman et al, 2003). The demand for identification of novel biomass degrading enzymes as well as for heterologous protein production at higher efficiency and reduced costs has catalyzed an interest in elucidating the genomic sequence of this fungus.…”