Detection and measurement of two-component systems that control dimorphism and virulence in fungi.
Systemic dimorphic fungi include six phylogenetically related ascomycetes. These organisms grow in a mold form in the soil on most continents around the world. After the mold spores, which are the infectious particles, are inhaled into the lung of a susceptible mammalian host, they undergo a morphological change into a pathogenic yeast form. The ability to convert to the yeast form is essential for this class of fungal agents to be pathogenic and produce disease. Temperature change is one key stimulus that triggers the phase transition from mold (25 degrees ) to yeast (37 degrees ). Genes that are expressed only in the pathogenic yeast form of these fungi have been identified to help explain how and why this phase transition is required for virulence. However, the regulators of yeast-phase specific genes, especially of phase transition from mold to yeast, have remained poorly understood. We used Agrobacterium-mediated gene transfer for insertional mutagenesis to create mutants that are defective in the phase transition and to identify genes that regulate this critical event. We discovered that a hybrid histidine kinase senses environmental signals such as temperature and regulates phase transition, dimorphism, and virulence in members of this fungal family. This chapter describes our approach to the identification and analysis of this global regulator.
Nemecek
Julie C
JC
Department of Pediatrics, University of Wisconsin-Madison School of Medicine and Public Health, Madison, Wisconsin, USA.
Wüthrich
Marcel
M
Klein
Bruce S
BS
D000595:Amino Acid Sequence; D005658:Fungi / Q000201:enzymology / Q000472:pathogenicity*; D018014:Gene Transfer Techniques; D000071677:Histidine Kinase; D008969:Molecular Sequence Data; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics; D034622:RNA Interference; D012098:Reproduction / Q000235:genetics; D012231:Rhizobium / Q000235:genetics; D014774:Virulence / Q000235:genetics*
D011494:Protein Kinases; D000071677:Histidine Kinase