Gain in cellular capacity to manage oxidative stress is associated with increased lifespan, and failure results in cellular damage triggering diseases such as cancer and diabetes. Though the cell has conserved pathways to respond to oxidative stress, much remains to be discovered. Our lab is interested in SKN-1, a major transcription factor that regulates the phase II detoxification response to oxidative stress. However, we recently discovered that SKN-1 is important for the activation and repression of a number of genes that influence autophagy and ER functions both during stress and in the absence of stress. This exciting preliminary data suggest that SKN-1 potentially aids in maintenance of cellular stability, or cellular homeostasis. I will be investigating the mechanism of SKN-1 regulation and whether a direct connection exists between Endoplasmic Reticulum (ER) and autophagy function during homeostasis and oxidative stress. In order to study stress in the context of a whole organism, I will be using the model organism C. elegans, which can be genetically manipulated and analyzed quickly in a high throughput manner at low cost. Understanding how SKN-1 restores the organism after stress and regulates stress to maintain homeostasis might uncover potential therapeutic targets for disease and disease prevention.
Specific Aim 1 : Does SKN-1 aid in cellular homeostasis by regulating autophagy and ER function? 1 will investigate whether SKN-1 also directs transcription of autophagy and ER stress genes by probing whether SKN-1 binds to the predicted binding sites in the promoter regions in vivo. I will then determine if SKN-1 activity is necessary for autophagy and/or ER stress.
Specific Aim 2 : How is SKN-1 is regulated in the nucleus before and after stress? I will first determine whether SKN-1 can be found at the site of transcription and whether recruitment of SKN-1 depends on the acetylation state of chromatin. I will also address whether p300 or HDA-2 is still recruited in the absence of SKN-1 activity as well as whether SKN-1 directly interacts with HDA-2. If I find that a direct association between HDA-2 and SKN-1 exists, I will then investigate whether SKN-1 activity or recruitment affects genome stability.
Specific Aim 3 : What other novel factors/co-factors are required for recovery from oxidative stress? To further our understanding of the oxidative stress response, I will use the RNAi libraries for C. elegans from our lab to search genome-wide for additional factors that are necessary for the organism to efficiently recovery from oxidative stress.
|Glover-Cutter, Kira M; Lin, Stephanie; Blackwell, T Keith (2013) Integration of the unfolded protein and oxidative stress responses through SKN-1/Nrf. PLoS Genet 9:e1003701|