ArticleNature cell biology2016
Avoiding artefacts when counting polymerized actin in live cells with LifeAct fused to fluorescent proteins.
Article in Nature cell biology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 87 papers.
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Who cites it
87 citing papers in PubMed, 147 citations in OpenAlex.
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- A genetic strategy to allow detection of F-actin by phalloidin staining in diverse fungi.mSphere · 2025Article
- Cryo-EM structure revealed a novel F-actin binding motif in abioRxiv : the preprint server for biology · 2025Article
- Three types of actomyosin rings within a common cytoplasm exhibit distinct modes of contractility.bioRxiv : the preprint server for biology · 2025Article
- ADPact: A versatile sensor for ADP-F-actin.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Liquid-like condensates that bind actin promote assembly and bundling of actin filaments.Developmental cell · 2025Article
- Mechanisms of force transmission on cytokinesis Formin Cdc12 in fission yeast revealed by coiled-coil force sensors.bioRxiv : the preprint server for biology · 2025Article
- Structure of the F-tractin-F-actin complex.The Journal of cell biology · 2025Article
- Regulation of Yeast Cytokinesis by Calcium.Journal of fungi (Basel, Switzerland) · 2025Review
- Design and Use of AsLOV2-Based Optogenetic Tools for Actin Imaging.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Improved tools for live imaging of F-actin structures in yeast.Molecular biology of the cell · 2024Article
- The Mechanosensitive Pkd2 Channel Modulates the Recruitment of Myosin II and Actin to the Cytokinetic Contractile Ring.Journal of fungi (Basel, Switzerland) · 2024Article
- Liquid-like condensates that bind actin drive filament polymerization and bundling.bioRxiv : the preprint server for biology · 2024Article
- Tools for live-cell imaging of cytoskeletal and nuclear behavior in the unconventional yeast,Molecular biology of the cell · 2024Article
- Cooperative actin filament nucleation by the Arp2/3 complex and formins maintains the homeostatic cortical array in Arabidopsis epidermal cells.The Plant cell · 2024Article
27 more citing papers are in PubMed but not listed here.
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Authors and funding
3 authors at 1 institution in 1 country.
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Abstract
When tagged with a fluorescent protein, actin is not fully functional, so the LifeAct peptide fused to a fluorescent protein is widely used to localize actin filaments in live cells. However, we find that these fusion proteins have many concentration-dependent effects on actin assembly in vitro and in fission yeast cells. mEGFP-LifeAct inhibits actin assembly during endocytosis as well as assembly and constriction of the cytokinetic contractile ring. Purified mEGFP-LifeAct and LifeAct-mCherry bind actin filaments with Kd values of ∼10 μM. LifeAct-mCherry can promote actin filament nucleation and either promote or inhibit filament elongation. Both separately and together, profilin and formins suppress these effects. LifeAct-mCherry can also promote or inhibit actin filament severing by cofilin. These concentration-dependent effects mean that caution is necessary when overexpressing LifeAct fusion proteins to label actin filaments in cells. Therefore, we used low micromolar concentrations of tagged LifeAct to follow assembly and disassembly of actin filaments in cells. Careful titrations also gave an estimate of a peak of ∼190,000 actin molecules (∼500 μm) in the fission yeast contractile ring. These filaments shorten from ∼500 to ∼100 subunits as the ring constricts.
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