Cryptococcus neoformans is an environmental fungal pathogen that requires atmospheric levels of
oxygen for optimal growth. For the fungus to be able to establish an
infection, it must adapt to the low
oxygen concentrations in the host environment compared to its natural habitat. In order to investigate the
oxygen sensing mechanism in C. neoformans, we screened
T-DNA insertional mutants for
hypoxia-mimetic
cobalt chloride (CoCl(2))-sensitive mutants. All the CoCl(2)-sensitive mutants had a growth defect under low
oxygen conditions at 37 degrees C. The majority of mutants are compromised in their mitochondrial function, which is reflected by their reduced rate of respiration. Some of the mutants are also defective in mitochondrial membrane permeability, suggesting the importance of an intact respiratory system for survival under both high concentrations of
CoCl(2) as well as low
oxygen conditions. In addition, the mutants tend to accumulate intracellular
reactive oxygen species (ROS), and all mutants show sensitivity to various ROS generating chemicals. Gene expression analysis revealed the involvement of several pathways in response to
cobalt chloride. Our findings indicate
cobalt chloride sensitivity and/or sensitivity to low
oxygen conditions are linked to mitochondrial function,
sterol and
iron homeostasis, ubiquitination, and the ability of cells to respond to ROS. These findings imply that multiple pathways are involved in
oxygen sensing in C. neoformans.