Abstract
Background2,3-Butanedione monoxime (BDM) has been widely used as a non-muscle myosin inhibitor to investigate the role of non-muscle myosinII in the process of actin retrograde flow and other actin cytoskeletal processes. Recent reports show that BDM does not inhibit any non-muscle myosins so far tested, including nm-myosinII, prompting the question, how were these process affected in BDM studies?ResultsWe have found that treatment of mammalian cells with BDM for only 1 min blocks actin incorporation at the leading edge in a permeabilized cell system. We show that inhibition of actin incorporation occurs through de-localization of leading edge proteins involved in actin polymerization – the Arp2/3 complex, WAVE, and VASP – that de-localize concomitantly with the leading edge actin network.ConclusionDe-localization of actin leading edge components by BDM treatment is a newly described effect of this compound. It may explain many of the results previously ascribed to inhibition of non-muscle myosinII by BDM, particularly in studies of leading edge dynamics. Though this effect of BDM is intriguing, future studies probing actin dynamics at the leading edge should use more potent and specific inhibitors.
Highlights
The actin cytoskeleton is a dynamic system composed of actin filaments, proteins that regulate filament dynamics, and proteins that remodel and make use of the cytoskeletal network
An important aspect of actin cytoskeletal research has been the use of small molecules that affect actin dynamics and proteins that act on the cytoskeleton [1]. 2,3-butanedione monoxime (BDM) is a small molecule that inhibits the ATPase activity of muscle myosinII and reduces the force generated by this motor protein [2,3]
Arp3 is no longer localized at the leading edge with BDM treatment Since actin incorporation is not due to elongation of existing filaments and BDM does not affect pure actin dynamics in vitro, we examined the localization of Arp3, a subunit of the actin nucleation Arp2/3 complex
Summary
The actin cytoskeleton is a dynamic system composed of actin filaments, proteins that regulate filament dynamics, and proteins that remodel and make use of the cytoskeletal network. An important aspect of actin cytoskeletal research has been the use of small molecules that affect actin dynamics and proteins that act on the cytoskeleton [1]. 2,3-butanedione monoxime (BDM) is a small molecule that inhibits the ATPase activity of muscle myosinII and reduces the force generated by this motor protein [2,3]. BDM was introduced to the cell biology community as a millimolar inhibitor of multiple non-muscle myosins with greatest potency in inhibiting non-muscle myosinII [4]. It was subsequently used, in concert with other approaches, to study nm-myosinII function in a variety of biological processes [5,6,7,8]. Retrograde flow is thought to be (page number not for citation purposes)
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
