Available at: http://digitalcommons.calpoly.edu/theses/142
Date of Award
MS in Biological Sciences
Nikki L. Adams
The amount of solar ultraviolet radiation (UVR, 290-400 nm) reaching Earth’s surface is increasing due to ozone depletion and global climate change. Embryos of the purple sea urchin, Strongylocentrotus purpuratus, provide an ideal system for examining how UVR affects developing marine organisms and cells in general. To model the protein-mediated cell cycle response to UV-irradiation, six batches of S. purpuratus embryos were exposed to UVR, monitored for delays in the first mitotic division and examined for global proteomic changes. Embryos from each batch were exposed to or protected from artificial UVR for 25 or 60 min. Embryos treated with UVR for 60 min cleaved an average of 23.24 min (±1.92 s.e.m) later than the UV-protected embryos. Protein expression of UV-protected and UV-treated embryos was examined at 30 and 90 min post-fertilization using two dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D SDS-PAGE) and mass spectrometry (MS). Proteins were isoelectrically focused (pH 4-7) and separated by molecular weight using SDS-PAGE. At least 1,306 protein spots were detected in all gels. A total of 171 protein spots (13% of the detected proteome) migrated differently in UV-treated embryos at 30 min post-fertilization and 187 spots (14%) at 90 min post-fertilization (2-way ANOVA, P= 0.03, n=6). Our results identify the differential migration of proteins from multiple cellular pathways and are the first to indicate that the mechanisms involved in the protein mediated UV-induced developmental delay are integrated among pathways for cellular stress, protein turnover and translation, signal transduction, general metabolism and involve the cytoskeleton.