Cargando…

An Unbiased Analysis Method to Quantify mRNA Localization Reveals Its Correlation with Cell Motility

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 employe...

Descripción completa

Detalles Bibliográficos
Autores principales: Park, Hye Yoon, Trcek, Tatjana, Wells, Amber L., Chao, Jeffrey A., Singer, Robert H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4079260/
https://www.ncbi.nlm.nih.gov/pubmed/22832165
http://dx.doi.org/10.1016/j.celrep.2011.12.009
Descripción
Sumario: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 non-randomly 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.