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Title: Imaging of single mRNA molecules moving within a living cell nucleus

Journal Article · · Biochemical and Biophysical Research Communications
 [1];  [1];  [2];  [2];  [3]
  1. Major in Integrative Bioscience and Biomedical Engineering, Graduate School of Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 169-8555 (Japan)
  2. Department of Biological Science, Faculty of Science, Kumamoto University, 2-39-1, Kurokami, Kumamoto 860-8555 (Japan)
  3. Major in Integrative Bioscience and Biomedical Engineering, Graduate School of Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 169-8555 (Japan) and Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-0033 (Japan) and Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama 332-0012 (Japan)

In eukaryotic cells, pre-mRNAs are transcribed in the nucleus, processed by 5' capping, 3'-polyadenylation, and splicing, and exported to the cytoplasm for translation. To examine the nuclear mRNA transport mechanism, intron-deficient mRNAs of truncated {beta}-globin and EGFP were synthesized, fluorescently labeled in vitro, and injected into the nucleus of living Xenopus A6 cells. The trajectories of single mRNA molecules in the nucleus were visualized using video-rate confocal microscopy. Approximately half the mRNAs moved by Brownian motion in the nucleoplasm, except the nucleoli, with an apparent diffusion coefficient of 0.2 {mu}m{sup 2}/s, about 1/150 of that in water. The slow diffusion could not be explained by simple diffusion obeying the Stokes-Einstein equation, suggesting interactions of the mRNAs with nuclear components. The remaining mRNAs were stationary with an average residence time of about 30 s, comparable to the time required for mRNA diffusion from the site of synthesis to nuclear pores.

OSTI ID:
20798989
Journal Information:
Biochemical and Biophysical Research Communications, Vol. 344, Issue 3; Other Information: DOI: 10.1016/j.bbrc.2006.03.202; PII: S0006-291X(06)00775-3; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
Country of Publication:
United States
Language:
English