Real-time visualization of ZBP1 association with β-actin mRNA during transcription and localization

Yuri Oleynikov, Robert H. Singer

Research output: Contribution to journalArticle

146 Citations (Scopus)

Abstract

Background: mRNA localization in somatic cells is an important mechanism for gene expression regulation. In fibroblasts, the protein ZBP1 associates with the sequence that localizes β-actin mRNA to the leading edge of fibroblasts, augmenting motility. β-actin mRNA localizes in a cytoskeleton-dependent manner, depending on intact actin and myosin ATP-hydrolysis, and is largely bound to the actin cytoskeleton. The ZBP1 protein contains four KH RNA binding domains and a classic RBD RNA binding domain. It also contains a putative nuclear import and export sequence, suggesting a nuclear phase in this protein's function. Results: Using high-speed imaging, we show here the targeting of this RNA binding protein to β-actin pre-mRNA transcripts in the nuclei of living cells and measure the residence time of the RNA-protein complex before it leaves the transcription site. Then, the RNA-protein particle is exported to the cytoplasm, where it localizes at velocities of 0.6 μm/s by using actin filaments and/or microtubules. This RNA-ZBP1 complex is required for cytoplasmic localization in fibroblasts; mislocalizing the protein also mislocalizes the RNA, and expressing the protein in a ZBP1-deficient cell line induces β-actin mRNA localization. Conclusions: This work demonstrates that the RNA-protein association, essential for cytoplasmic localization, begins as soon as the RNA is transcribed. The ZBP1 then forms a ribonucleoprotein particle and moves in a myosin-dependent fashion by using the cytoskeleton for directional transport.

Original languageEnglish (US)
Pages (from-to)199-207
Number of pages9
JournalCurrent Biology
Volume13
Issue number3
DOIs
StatePublished - Feb 2003

Fingerprint

Transcription
actin
Actins
Visualization
transcription (genetics)
Association reactions
RNA
Messenger RNA
Proteins
Fibroblasts
proteins
fibroblasts
Cell Nucleus Active Transport
Myosins
myosin
cytoskeleton
microfilaments
Cytoskeleton
Actin Cytoskeleton
Gene expression regulation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)

Cite this

Real-time visualization of ZBP1 association with β-actin mRNA during transcription and localization. / Oleynikov, Yuri; Singer, Robert H.

In: Current Biology, Vol. 13, No. 3, 02.2003, p. 199-207.

Research output: Contribution to journalArticle

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