Regulation Of Pro Longevity Ros By Ros Handling Enzymes In C Elegans PDF Download

"Reactive oxygen species (ROS) are highly reactive molecules which have the potential to damage all biological macromolecules, yet they can also function as signalling molecules (Shibata, Branicky et al. 2003, Sena and Chandel 2012, Putker, Madl et al. 2013, Holmstrom and Finkel 2014). In this thesis, I explore the regulatory effects of reactive oxygen species (ROS) on the Ras signalling cascade in Caenorhabditis elegans vulva development and aging. Oncogenic Ras is one of the deadliest drivers of human cancer (Papke and Der 2017). 30% of all human cancers are driven by oncogenic Ras mutations, including a high percentage of the most lethal cancers: lung, colon and pancreatic cancer (Cox, Fesik et al. 2014). Unfortunately, despite more than three decades of research, no effective pharmacological inhibitor of oncogenic Ras has yet been clinically approved (McCormick 2015). As such, there remains a critical need for innovative therapeutic strategies to counteract the effects of oncogenic Ras mutations. In C. elegans, let-60 encodes the homolog of mammalian Ras isoforms. This gene regulates many processes including the development of the vulva (Reiner and Lundquist 2016). We used the let-60(n1046gf) mutant which encodes an oncogenic LET-60 protein (LET-60rasgf) and exhibits a multivulva (Muv) phenotype (Han and Sternberg 1990) to study regulatory effects of ROS on LET-60rasgf signalling. We uncovered a ROS signalling network which regulates vulval formation through LET-60rasgf signalling and by other mechanisms. In particular, we found that oxidation of the conserved cysteine 118 (C118) of LET-60rasgf by cytoplasmic hydrogen peroxide inhibits its activity and leads to the inhibition of the Muv phenotype. Confirming the effectiveness of hydrogen peroxide to inhibit oncogenic LET-60rasgf, we found that substituting C118 with an aspartic acid (C118D), which mimics permanent oxidation by hydrogen peroxide, fully suppressed LET-60rasgf. Increased production of mitochondrial ROS (mtROS) increases the lifespan of C. elegans (Van Raamsdonk and Hekimi 2009, Yang and Hekimi 2010). However, the exact molecular targets of mtROS that trigger longevity are unknown. In our study, we also found that C118 of LET-60 appears to be a target of pro-longevity mtROS. Indeed, in some situations, the pro-longevity action of mtROS is inhibited by the substitution of C118 with a non-oxidable serine (C118S), suggesting that C118 is indeed one of the targets of pro-longevity mtROS"--