The Effect of ELCI on Cleaving After DNA Damage (2021)

In eukaryotes when DNA is transcribed, pre-mRNA is made by RNA polymerase. It is then cleaved, and a tail structure added before the mRNA is transported to the cytoplasm to be translated into protein. Specific protein complexes called cleavage and polyadenylation factors are required for the cleavage and tail addition process. This process of tail addition is impaired following certain types of DNA damage. ElcI is a protein degradation factor. Our hypothesis is that Elc1 binds to cleavage and polyadenylation factors to degrade these proteins following DNA damage. We use a process called immunoprecipitation to test this hypothesis. We use extracts of a specific strain of yeast with a tag on ElcI that will bind to an antibody conjugated bead. We then test the proteins bound to Elc1 by analyzing this bead- antibody -Elc1 complex in order to check if cleavage and polyadenylation factors bind to Elc1 following DNA damage.