title: Molecular detection and characterisation of fungal heat shock protein 60.
abstract: Heat shock proteins (Hsp) are highly conserved molecules, which are both constitutively expressed and up-regulated in response to various stress conditions. In particular, fungal Hsp60 can act as immunodominant antigens and facilitate powerful immunological properties. A possible cellular heat shock response was investigated in eight fungi (Aspergillus fumigatus, Aspergillus terreus, Penicillium chrysogenum, Cladosporium cladosporioides, Scedosporium apiospermum, Trichophyton mentagrophytes, Candida albicans and Saccharomyces cerevisiae). Fully automated RNA extraction was followed by quantitative real-time RT-PCR targeting fungus-specific Hsp60 mRNA and sequencing of the amplicon. Levels of temperature-dependent gene expression were evaluated and rates of similarity and identity were compared. While Hsp60 mRNA was constitutively expressed in all the samples tested, a temperature-dependent induction was not shown in C. cladosporioides. In the 80-amino acid fragment from the hypothetical protein, 66% of the amino acids were identical, 20% showed a conserved and 8% a semi-conserved substitution. Our findings should contribute to a better understanding of host-pathogen relationship and suggest that fungal Hsp60 under temperature-related stress conditions might act as an immunogenic trigger in orchestrating fungi-related diseases.
authors.affiliation: Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Austria.
authors.forename: Reinhard B
authors.initials: RB
authors.lastname: Raggam
authors.forename: Helmut J F
authors.initials: HJ
authors.lastname: Salzer
authors.forename: Egon
authors.initials: E
authors.lastname: Marth
authors.forename: Bettina
authors.initials: B
authors.lastname: Heiling
authors.forename: Astrid H
authors.initials: AH
authors.lastname: Paulitsch
authors.forename: Walter
authors.initials: W
authors.lastname: Buzina
mesh_terms: D019943:Amino Acid Substitution; D018834:Chaperonin 60 / Q000235:genetics* / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D005658:Fungi / Q000528:radiation effects*; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D006358:Hot Temperature; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013312:Stress, Physiological
chemical_list: D018834:Chaperonin 60; D005656:Fungal Proteins