ArticleBMC microbiology2011
Calcium/calmodulin kinase1 and its relation to thermotolerance and HSP90 in Sporothrix schenckii: an RNAi and yeast two-hybrid study.
Article in BMC microbiology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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24 citing papers in PubMed, 45 citations in OpenAlex.
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- Disruption of protein rhamnosylation affects theCell surface (Amsterdam, Netherlands) · 2021Article
- Transcriptome Analysis of Dimorphic Fungus Sporothrix schenckii Exposed to Temperature Stress.International microbiology : the official journal of the Spanish Society for Microbiology · 2021Article
- Identification and Phenotypic Characterization of Hsp90 Phosphorylation Sites That Modulate Virulence Traits in the Major Human Fungal PathogenFrontiers in cellular and infection microbiology · 2021Article
- Advances in Molecular Tools and In Vivo Models for the Study of Human Fungal Pathogenesis.Microorganisms · 2020Review
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- SEM and qRT-PCR revealed quercetin inhibits morphogenesis ofMycology · 2020Article
- Ste20 is crucial for dimorphic switching of sporothrix schenckii and affects its global transcriptome.International journal of clinical and experimental pathology · 2020Article
- Silencing ofInfection and drug resistance · 2019Article
- TheFrontiers in microbiology · 2017Article
- Morphology Changes in Human Fungal Pathogens upon Interaction with the Host.Journal of fungi (Basel, Switzerland) · 2017Article
- Heat Stress Modulates Mycelium Growth, Heat Shock Protein Expression, Ganoderic Acid Biosynthesis, and Hyphal Branching of Ganoderma lucidum via Cytosolic Ca2.Applied and environmental microbiology · 2016Article
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5 authors at 1 institution in 1 country.
Funding
Abstract
backgroundSporothrix schenckii is a pathogenic dimorphic fungus of worldwide distribution. It grows in the saprophytic form with hyaline, regularly septated hyphae and pyriform conidia at 25°C and as the yeast or parasitic form at 35°C. Previously, we characterized a calcium/calmodulin kinase in this fungus. Inhibitors of this kinase were observed to inhibit the yeast cell cycle in S. schenckii.
resultsThe presence of RNA interference (RNAi) mechanism in this fungus was confirmed by the identification of a Dicer-1 homologue in S. schenckii DNA. RNAi technology was used to corroborate the role of calcium/calmodulin kinase I in S. schenckii dimorphism. Yeast cells were transformed with the pSilent-Dual2G (pSD2G) plasmid w/wo inserts of the coding region of the calcium/calmodulin kinase I (sscmk1) gene. Transformants were selected at 35°C using resistance to geneticin. Following transfer to liquid medium at 35°C, RNAi transformants developed as abnormal mycelium clumps and not as yeast cells as would be expected. The level of sscmk1 gene expression in RNAi transformants at 35°C was less than that of cells transformed with the empty pSD2G at this same temperature. Yeast two-hybrid analysis of proteins that interact with SSCMK1 identified a homologue of heat shock protein 90 (HSP90) as interacting with this kinase. Growth of the fungus similar to that of the RNAi transformants was observed in medium with geldanamycin (GdA, 10 μM), an inhibitor of HSP90.
conclusionsUsing the RNAi technology we silenced the expression of sscmk1 gene in this fungus. RNAi transformants were unable to grow as yeast cells at 35°C showing decreased tolerance to this temperature. The interaction of SSCMK1 with HSP90, observed using the yeast two-hybrid assay suggests that this kinase is involved in thermotolerance through its interaction with HSP90. SSCMK1 interacted with the C terminal domain of HSP90 where effector proteins and co-chaperones interact. These results confirmed SSCMK1 as an important enzyme involved in the dimorphism of S. schenckii, necessary for the development of the yeast phase of this fungus. Also this study constitutes the first report of the transformation of S. schenckii and the use of RNAi to study gene function in this fungus.
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