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

Hye Yoon Park; Tatjana Trcek; Amber L. Wells; Jeffrey A. Chao; Robert H. Singer

doi:10.1016/j.celrep.2011.12.009

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

Hye Yoon Park, Tatjana Trcek, Amber L. Wells, Jeffrey A. Chao, Robert H. Singer

Cell Biology

Research output: Contribution to journal › Article › peer-review

55 Scopus citations

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.

Original language	English (US)
Pages (from-to)	179-184
Number of pages	6
Journal	Cell Reports
Volume	1
Issue number	2
DOIs	https://doi.org/10.1016/j.celrep.2011.12.009
State	Published - Feb 23 2012

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/j.celrep.2011.12.009

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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.",

author = "Park, {Hye Yoon} and Tatjana Trcek and Wells, {Amber L.} and Chao, {Jeffrey A.} and Singer, {Robert H.}",

note = "Funding Information: Microscopy equipment for the live cell imaging experiments was provided by the Gruss Lipper Biophotonics Center. This work was supported by NIH GM57071, GM84364 and GM86217 to R.H.S. H.Y.P. was supported by National Research Service Awards F32-GM087122. ",

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AU - Park, Hye Yoon

AU - Trcek, Tatjana

AU - Wells, Amber L.

AU - Chao, Jeffrey A.

AU - Singer, Robert H.

N1 - Funding Information: Microscopy equipment for the live cell imaging experiments was provided by the Gruss Lipper Biophotonics Center. This work was supported by NIH GM57071, GM84364 and GM86217 to R.H.S. H.Y.P. was supported by National Research Service Awards F32-GM087122.

PY - 2012/2/23

Y1 - 2012/2/23

N2 - 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.

AB - 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.

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

Abstract

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