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Nanoscale dynamics and localization of single endogenous mRNAs in stress granules.
Sugawara, Ko; Uno, Shin-Nosuke; Kamiya, Mako; Sakamoto, Akihiko; Urano, Yasuteru; Funatsu, Takashi; Okabe, Kohki.
Afiliação
  • Sugawara K; Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan.
  • Uno SN; RIKEN Center for Biosystems Dynamics Research, Hyogo 650-0047, Japan.
  • Kamiya M; Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
  • Sakamoto A; Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan.
  • Urano Y; Department of Life Science and Technology, Tokyo Institute of Technology, Kanagawa 226-8501, Japan.
  • Funatsu T; Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan.
  • Okabe K; Department of Pharmacology, Yamaguchi University Graduate School of Medicine, Yamaguchi 755-8505, Japan.
Nucleic Acids Res ; 2024 Jul 29.
Article em En | MEDLINE | ID: mdl-39069641
ABSTRACT
Stress granules (SGs) are cytoplasmic messenger ribonucleoprotein granules transiently formed in stressed mammalian cells. Although SG components have been well characterized, detailed insights into the molecular behavior inside SGs remain unresolved. We investigated nanoscale dynamics and localization of endogenous mRNAs in SGs combining single mRNA tracking and super-resolution localization microscopy. First, we developed a methodology for tracking single mRNAs within SGs, revealing that although mRNAs in SGs are mainly stationary (∼40%), they also move in a confined (∼25%) or freely diffusing (∼35%) manner. Second, the super-resolution localization microscopy showed that the mRNAs in SGs are heterogeneously distributed and partially form high-density clusters. Third, we simultaneously performed single mRNA tracking and super-resolution microscopy in SGs, demonstrating that single mRNA trajectories are mainly found around high-density clusters. Finally, a quantitative analysis of mRNA localization and dynamics during stress removal was conducted using live super-resolution imaging and single-molecule tracking. These results suggest that SGs have a highly organized structure that enables dynamic regulation of the mRNAs at the nanoscale, which is responsible for the ordered formation and the wide variety of functions of SGs.

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Nucleic Acids Res Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Japão

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Nucleic Acids Res Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Japão