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Title: Single-molecule imaging of {beta}-actin mRNAs in the cytoplasm of a living cell

Abstract

{beta}-Actin mRNA labeled with an MS2-EGFP fusion protein was expressed in chicken embryo fibroblasts and its localization and movement were analyzed by single-molecule imaging. Most {beta}-Actin mRNAs localized to the leading edge, while some others were observed in the perinuclear region. Singe-molecule tracking of individual mRNAs revealed that the majority of mRNAs were in unrestricted Brownian motion at the leading edge and in restricted Brownian motion in the perinuclear region. The macroscopic diffusion coefficient of mRNA (D{sub MACRO}) at the leading edge was 0.3 {mu}m{sup 2}/s. On the other hand, D{sub MACRO} in the perinuclear region was 0.02 {mu}m{sup 2}/s. The destruction of microfilaments with cytochalasin D, which is known to delocalize {beta}-actin mRNAs, led to an increase in D{sub MACRO} to 0.2 {mu}m{sup 2}/s in the perinuclear region. These results suggest that the microstructure, composed of microfilaments, serves as a barrier for the movement of {beta}-actin mRNA.

Authors:
;  [1];  [2];  [3];  [1];  [4]
  1. Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo-ku, Tokyo 113-0033 (Japan)
  2. Laboratory of Genome Technology, Department of Human Genome Research, Kazusa DNA Research Institute, Kisarazu, Chiba 292-0818 (Japan)
  3. Major in Integrative Bioscience and Biomedical Engineering, Graduate School of Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 169-8555 (Japan)
  4. (Japan)
Publication Date:
OSTI Identifier:
22209811
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; Journal Volume: 315; Journal Issue: 7; Other Information: Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACTIN; BROWNIAN MOVEMENT; CHICKENS; CYTOPLASM; EMBRYOS; FIBROBLASTS; FILAMENTS; MESSENGER-RNA; MICROSTRUCTURE; MOLECULES

Citation Formats

Yamagishi, Mai, Ishihama, Yo, Shirasaki, Yoshitaka, Kurama, Hideki, Funatsu, Takashi, E-mail: funatsu@mail.ecc.u-tokyo.ac.jp, and Center for NanoBio Integration, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656. Single-molecule imaging of {beta}-actin mRNAs in the cytoplasm of a living cell. United States: N. p., 2009. Web. doi:10.1016/J.YEXCR.2009.02.009.
Yamagishi, Mai, Ishihama, Yo, Shirasaki, Yoshitaka, Kurama, Hideki, Funatsu, Takashi, E-mail: funatsu@mail.ecc.u-tokyo.ac.jp, & Center for NanoBio Integration, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656. Single-molecule imaging of {beta}-actin mRNAs in the cytoplasm of a living cell. United States. doi:10.1016/J.YEXCR.2009.02.009.
Yamagishi, Mai, Ishihama, Yo, Shirasaki, Yoshitaka, Kurama, Hideki, Funatsu, Takashi, E-mail: funatsu@mail.ecc.u-tokyo.ac.jp, and Center for NanoBio Integration, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656. Wed . "Single-molecule imaging of {beta}-actin mRNAs in the cytoplasm of a living cell". United States. doi:10.1016/J.YEXCR.2009.02.009.
@article{osti_22209811,
title = {Single-molecule imaging of {beta}-actin mRNAs in the cytoplasm of a living cell},
author = {Yamagishi, Mai and Ishihama, Yo and Shirasaki, Yoshitaka and Kurama, Hideki and Funatsu, Takashi, E-mail: funatsu@mail.ecc.u-tokyo.ac.jp and Center for NanoBio Integration, The University of Tokyo, Bunkyo-ku, Tokyo 113-8656},
abstractNote = {{beta}-Actin mRNA labeled with an MS2-EGFP fusion protein was expressed in chicken embryo fibroblasts and its localization and movement were analyzed by single-molecule imaging. Most {beta}-Actin mRNAs localized to the leading edge, while some others were observed in the perinuclear region. Singe-molecule tracking of individual mRNAs revealed that the majority of mRNAs were in unrestricted Brownian motion at the leading edge and in restricted Brownian motion in the perinuclear region. The macroscopic diffusion coefficient of mRNA (D{sub MACRO}) at the leading edge was 0.3 {mu}m{sup 2}/s. On the other hand, D{sub MACRO} in the perinuclear region was 0.02 {mu}m{sup 2}/s. The destruction of microfilaments with cytochalasin D, which is known to delocalize {beta}-actin mRNAs, led to an increase in D{sub MACRO} to 0.2 {mu}m{sup 2}/s in the perinuclear region. These results suggest that the microstructure, composed of microfilaments, serves as a barrier for the movement of {beta}-actin mRNA.},
doi = {10.1016/J.YEXCR.2009.02.009},
journal = {Experimental Cell Research},
number = 7,
volume = 315,
place = {United States},
year = {Wed Apr 15 00:00:00 EDT 2009},
month = {Wed Apr 15 00:00:00 EDT 2009}
}
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