RESUMO
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.
Assuntos
Chaperonina 60/genética , Chaperonina 60/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Fungos/efeitos da radiação , Regulação Fúngica da Expressão Gênica , Estresse Fisiológico , Substituição de Aminoácidos , Perfilação da Expressão Gênica , Temperatura Alta , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosAssuntos
Abscesso Encefálico/microbiologia , Diabetes Mellitus/patologia , Sinusite Frontal/microbiologia , Schizophyllum/isolamento & purificação , Anfotericina B/farmacologia , Anfotericina B/uso terapêutico , Antifúngicos/farmacologia , Antifúngicos/uso terapêutico , Encéfalo/diagnóstico por imagem , Encéfalo/microbiologia , Abscesso Encefálico/tratamento farmacológico , DNA Fúngico/genética , Sinusite Frontal/cirurgia , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Micoses/diagnóstico , Micoses/tratamento farmacológico , Micoses/microbiologia , Radiografia , Schizophyllum/efeitos dos fármacos , Schizophyllum/genéticaRESUMO
Heat shock proteins or chaperones are found in mitochondrial and cytosolic compartments of cells. They are responsible for the correct folding of proteins and are up-regulated in reaction to various stressors. In addition, when released or presented on the surface of cells, they may play an important role in inflammatory and immunomodulating processes. To identify and characterize hsp60 in the common environmental mold Alternaria alternata, the fungus was cultivated and incubated at different temperatures to induce a possible heat shock response. Fully automated RNA extraction was followed by quantitative real-time RT-PCR targeting A. alternata specific Hsp60 mRNA and subsequent sequencing of the amplicon. While Hsp60 mRNA was constitutively expressed in all samples tested, a temperature-dependent expression of Hsp60 mRNA was observed. Sequencing revealed an identity of more than 85% to other fungal hsp60, indicating the existence of this protein in A. alternata.