title: An unbiased analysis method to quantify mRNA localization reveals its correlation with cell motility.
abstract: Localization of mRNA is a critical mechanism used by a large fraction of transcripts to restrict its translation to specific cellular regions. Although current high-resolution imaging techniques provide ample information, the analysis methods for localization have either been qualitative or employed quantification in nonrandomly selected regions of interest. Here, we describe an analytical method for objective quantification of mRNA localization using a combination of two characteristics of its molecular distribution, polarization and dispersion. The validity of the method is demonstrated using single-molecule FISH images of budding yeast and fibroblasts. Live-cell analysis of endogenous β-actin mRNA in mouse fibroblasts reveals that mRNA polarization has a half-life of ~16 min and is cross-correlated with directed cell migration. This novel approach provides insights into the dynamic regulation of mRNA localization and its physiological roles.
authors: Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors: Hye Yoon
authors: HY
authors: Park
authors: Tatjana
authors: T
authors: Trcek
authors: Amber L
authors: AL
authors: Wells
authors: Jeffrey A
authors: JA
authors: Chao
authors: Robert H
authors: RH
authors: Singer
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002465:Cell Movement / Q000235:genetics*; D002642:Chick Embryo; D003198:Computer Simulation; D004622:Embryo, Mammalian / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional / Q000379:methods*; D017403:In Situ Hybridization; D051379:Mice; D009010:Monte Carlo Method; D034443:RNA Transport / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism
chemical_list: D000199:Actins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins