An Analysis of Gene Expression During Exposure of Neurospora crassa Cells to Dual Stresses of High Temperature and 2-deoxyglucose
The model mycelial fungus Neurospora crassa shows a robust heat shock response that is protective. However, the wild-type strain is not resistant to heat shock when it is coupled with an inhibitory glucose analog. In contrast, mutation of a stress mitogen-activated kinase gene (os2) that is roughly orthologous to Saccharomyces cerevisiae hog1, renders these N. crassa cells resistant to the coupled stresses. With the University's Center for Mass Spectrometry and Proteomics, these researchers have conducted an iTRAQ experiment to compare the relative levels of proteins in the wild type and the os2 strains. This has revealed several intriguing differences, showing higher levels of heat shock proteins and mitochondrial proteins in the os2 strain. The researchers have found that protein synthesis is impaired in the dual stressed wild-type cells, during what is usually the recovery phase of heat shock. However, the os2 strain does recover normal protein synthesis during the dual stress. The researchers have performed preliminary Northern experiments that indicate that there are likely differences between these strains at the RNA level for some key proteins. They have performed an RNA-seq experiment with wild type and os2 RNA samples, purified from wild type and os2 cells at normal temperature, heat shock temperature, and heat shock coupled with glucose analog. They have analyzed these data and are combining them with the iTRAQ data. They are continuing to use MSI to verify their analyses.
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