Dataset Information

Dynamics of native ?-actin mRNA transport in the cytoplasm.

ABSTRACT: Transport of messenger RNAs (mRNAs) in the cytoplasm is essential for localization to translation sites and for post-transcriptional regulation. Utilizing single-RNA sensitive probes and real-time fluorescence microscopy, we accurately quantified the dynamics of native, non-engineered, ?-actin mRNAs within the cytoplasm of epithelial cells and fibroblasts for the first time. Using single-particle tracking and temporal analysis, we determined that native ?-actin mRNAs, under physiologic conditions, exhibit bursts of intermittent, processive motion on microtubules, interspersed between time periods of diffusive motion, characterized by non-thermal enhanced diffusivity. When transport processes were perturbed via ATP depletion, temperature reduction, dynamitin overexpression and chemical inhibitors, processive motion was diminished or eliminated and diffusivity was reduced. These data support a model whereby processive, motor-driven motion is responsible for long-distance mRNA transport.

SUBMITTER: Lifland AW

PROVIDER: S-EPMC3134163 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Dynamics of native β-actin mRNA transport in the cytoplasm.

Lifland Aaron W AW Zurla Chiara C Yu Joana J Santangelo Philip J PJ

Traffic (Copenhagen, Denmark) 20110518 8

Transport of messenger RNAs (mRNAs) in the cytoplasm is essential for localization to translation sites and for post-transcriptional regulation. Utilizing single-RNA sensitive probes and real-time fluorescence microscopy, we accurately quantified the dynamics of native, non-engineered, β-actin mRNAs within the cytoplasm of epithelial cells and fibroblasts for the first time. Using single-particle tracking and temporal analysis, we determined that native β-actin mRNAs, under physiologic condition ...[more]

PMID: 21518164

Similar Datasets

Project description:Guidance cues trigger fast responses in axonal growth cones such as directional turning and collapse that require local protein synthesis. An attractive cue-gradient, such as Netrin-1, triggers de novo synthesis of β-actin localized to the near-side compartment of the growth cone that promotes F-actin assembly and attractive steering. How this precise spatial asymmetry in mRNA translation arises across the small expanse of the growth cone is poorly understood. Pre-localized mRNAs in the vicinity of activated receptors could be selectively translated and/or new mRNAs could be trafficked into the area. Here we have performed live imaging of fluorescent-tagged β-actin mRNA to investigate mRNA trafficking dynamics in Xenopus retinal ganglion cell (RGC) axons and growth cones in response to Netrin-1. A Netrin-1 gradient was found to elicit the transport of β-actin mRNA granules to the near-side of growth cones within a 4-7 min window. This polarized mRNA trafficking depended on the 3' untranslated region (UTR) since mRNA-Δ3'UTR mutant failed to exhibit cue-induced localization. Global application of Netrin-1 significantly increased the anterograde movement of β-actin mRNA along axons and also promoted microtubule-dependent mRNA excursions from the central domain of the growth cone into the periphery (filopodia and lamellipodia). Dual channel imaging revealed β-actin mRNA riding behind the microtubule plus-end tracking protein, EB1, in movements along dynamic microtubules into filopodia. The mRNA-EB1 movements were unchanged by a Netrin-1 gradient indicating the dynamic microtubules themselves do not underlie the cue-induced polarity of RNA movement. Finally, fast-moving elongated "worm-like" trains of Cy3-RNA, distinct from mitochondria, were seen transporting RNA along axons in vitro and in vivo suggesting the existence of a novel transport organelle. Overall, the results provide evidence that the axonal trafficking of β-actin mRNA can be regulated by the guidance cue Netrin-1 to transduce the polarity of an extracellular stimulus and that the 3'UTR is essential for this cue-induced regulation.

Dataset Information

Dynamics of native ?-actin mRNA transport in the cytoplasm.

Publications

Dynamics of native β-actin mRNA transport in the cytoplasm.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets