11a. Two papers covered in class related to the question of how common mRNA localization is in “everyday” cells, i.e. cells other than eggs or embryos, the Mingle et. al. (2005) and the Mili et. al. (2008) papers. For Mingle et al. (2005), provide a brief summary of the methods, the results and their conclusions. Be sure to include information regarding which specific elements of the cytoskeleton were implicated in the localization of the mRNAs in question.
- Use FISH-TSA to localize mRNA for b-actin and Arp2 and Arp3: All seven Arp2/3 complex mRNAs localize to human foreskin fibroblasts.
- Looked at pattern of dispersion in Poly-A RNAs: shows disperse hybridization pattern
- Looked at localization of Chick Arp2 with and without serum, to see if serum impacted localization: Arp2 with serum showed increased localization over time, and without serum vice versa.
- Looked at WHAT component of the cytoskeleton is necessary for localization, using Cytochalasin D (actin poly inhib) and Colchicine (mt poly inhib): Showed localization of Arp2/3 mRNA dependent on MF and MTs, wherease localization of b-actin mRNA dependent on MFilaments only
- Conclusions: 1. mRNA encoding proteins that function together co-localize together. 2. Localization dependent on intact cytoskeleton, particularly with MF and MTs intact