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PMID: 21880897 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

A pharmacological cocktail for arresting actin dynamics in living cells.

Molecular biology of the cell ·Vol. 22 ·No. 21 ·2011-11-00 ·Pages 3986-94

Peng GE, Wilson SR, Weiner OD

Abstract

The actin cytoskeleton is regulated by factors that influence polymer assembly, disassembly, and network rearrangement. Drugs that inhibit these events have been used to test the role of actin dynamics in a wide range of cellular processes. Previous methods of arresting actin rearrangements take minutes to act and work well in some contexts, but can lead to significant actin reorganization in cells with rapid actin dynamics, such as neutrophils. In this paper, we report a pharmacological cocktail that not only arrests actin dynamics but also preserves the structure of the existing actin network in neutrophil-like HL-60 cells, human fibrosarcoma HT1080 cells, and mouse NIH 3T3 fibroblast cells. Our cocktail induces an arrest of actin dynamics that initiates within seconds and persists for longer than 10 min, during which time cells maintain their responsivity to external stimuli. With this cocktail, we demonstrate that actin dynamics, and not simply morphological polarity or actin accumulation at the leading edge, are required for the spatial persistence of Rac activation in HL-60 cells. Our drug combination preserves the structure of the existing cytoskeleton while blocking actin assembly, disassembly, and rearrangement, and should prove useful for investigating the role of actin dynamics in a wide range of cellular signaling contexts.

MeSH Terms
Actins/antagonists & inhibitors,metabolism Amides/pharmacology Animals Bridged Bicyclo Compounds, Heterocyclic/pharmacology Cell Polarity Cell Shape Depsipeptides/pharmacology Enzyme Activation Fluorescence Recovery After Photobleaching HL-60 Cells Humans Mice Microscopy, Fluorescence NIH 3T3 Cells Protein Multimerization/drug effects Pyridines/pharmacology Thiazolidines/pharmacology Time-Lapse Imaging rac GTP-Binding Proteins/metabolism
Chemicals
Actins Amides Bridged Bicyclo Compounds, Heterocyclic Depsipeptides Pyridines Thiazolidines jasplakinolide Y 27632 rac GTP-Binding Proteins latrunculin B
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Peng Grace E
Department of Biochemistry and Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA 94158, USA.
Wilson Sarah R
Weiner Orion D
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.E11-04-0379
Published
2011-11-00
Epub
2011-00-31
Pages
3986-94
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC3204061
Subset
IM
Grants
NIGMS NIH HHS · R01 GM084040 · United States
NIGMS NIH HHS · R01 GM084040-04 · United States
NIGMS NIH HHS · R01-GM084040 · United States
External Links
PubMed source DOI: 10.1091/mbc.E11-04-0379 PMC Full Text View journal details
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