Arp2/3 complex inhibitors adversely affect actin cytoskeleton remodeling in the cultured murine kidney collecting duct M-1 cells

Daria V. Ilatovskaya; Vladislav Chubinskiy-Nadezhdin; Tengis S. Pavlov; Leonid S. Shuyskiy; Viktor Tomilin; Oleg Palygin; Alexander Staruschenko; Yuri A. Negulyaev

doi:10.1007/s00441-013-1710-y

Arp2/3 complex inhibitors adversely affect actin cytoskeleton remodeling in the cultured murine kidney collecting duct M-1 cells

Daria V. Ilatovskaya, Vladislav Chubinskiy-Nadezhdin, Tengis S. Pavlov, Leonid S. Shuyskiy, Viktor Tomilin, Oleg Palygin, Alexander Staruschenko, Yuri A. Negulyaev

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

Abstract

Dynamic remodeling of the actin cytoskeleton plays an essential role in cell migration and various signaling processes in living cells. One of the critical factors that controls the nucleation of new actin filaments in eukaryotic cells is the actin-related protein 2/3 (Arp2/3) complex. Recently, two novel classes of small molecules that bind to different sites on the Arp2/3 complex and inhibit its ability to nucleate F-actin have been discovered and described. The current study aims at investigating the effects of CK-0944666 (CK-666) and its analogs (CK-869 and inactive CK-689) on the reorganization of the actin microfilaments in the cortical collecting duct cell line, M-1. We show that treatment with CK-666 and CK869 results in the reorganization of F-actin and drastically affects cell motility rate. The concentrations of the compounds used in this study (100-200 μM) neither cause loss of cell viability nor influence cell shape or monolayer integrity; hence, the effects of described compounds were not due to structural side effects. Therefore, we conclude that the Arp2/3 complex plays an important role in cell motility and F-actin reorganization in M-1 cells. Furthermore, CK-666 and its analogs are useful tools for the investigation of the Arp2/3 complex.

Original language	English (US)
Pages (from-to)	783-792
Number of pages	10
Journal	Cell and Tissue Research
Volume	354
Issue number	3
DOIs	https://doi.org/10.1007/s00441-013-1710-y
State	Published - Dec 2013
Externally published	Yes

Keywords

Actin filaments
Arp2/3 complex
CK-0944666
Cell motility

ASJC Scopus subject areas

Pathology and Forensic Medicine
Histology
Cell Biology

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10.1007/s00441-013-1710-y

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@article{aee2e621f7f54f7e8ee9d1ccf8dd4e54,

title = "Arp2/3 complex inhibitors adversely affect actin cytoskeleton remodeling in the cultured murine kidney collecting duct M-1 cells",

abstract = "Dynamic remodeling of the actin cytoskeleton plays an essential role in cell migration and various signaling processes in living cells. One of the critical factors that controls the nucleation of new actin filaments in eukaryotic cells is the actin-related protein 2/3 (Arp2/3) complex. Recently, two novel classes of small molecules that bind to different sites on the Arp2/3 complex and inhibit its ability to nucleate F-actin have been discovered and described. The current study aims at investigating the effects of CK-0944666 (CK-666) and its analogs (CK-869 and inactive CK-689) on the reorganization of the actin microfilaments in the cortical collecting duct cell line, M-1. We show that treatment with CK-666 and CK869 results in the reorganization of F-actin and drastically affects cell motility rate. The concentrations of the compounds used in this study (100-200 μM) neither cause loss of cell viability nor influence cell shape or monolayer integrity; hence, the effects of described compounds were not due to structural side effects. Therefore, we conclude that the Arp2/3 complex plays an important role in cell motility and F-actin reorganization in M-1 cells. Furthermore, CK-666 and its analogs are useful tools for the investigation of the Arp2/3 complex.",

keywords = "Actin filaments, Arp2/3 complex, CK-0944666, Cell motility",

author = "Ilatovskaya, {Daria V.} and Vladislav Chubinskiy-Nadezhdin and Pavlov, {Tengis S.} and Shuyskiy, {Leonid S.} and Viktor Tomilin and Oleg Palygin and Alexander Staruschenko and Negulyaev, {Yuri A.}",

note = "Funding Information: Acknowledgments The authors would like to acknowledge Dr. Grigoriy Stein (Institute of Cytology RAS) for help with microscopy experiments and sincerely thank Vladislav Levchenko (Medical College of Wisconsin) for help with the MTT cytotoxicity assay and critical reading of the manuscript. We also acknowledge the help of Glenn Slocum and Bradley Endres (Medical College of Wisconsin) for helpful discussion and correction of the manuscript. The laboratory of Dr. A.N. Tomilin (Institute of Cytology RAS, St. Petersburg) is recognized for sharing the antibodies for immunofluorescence. This research was supported by R01HL108880 from the National Heart, Lung, and Blood Institute and the American Diabetes Association grant 1-10-BS-168 (to A.S.), Russian Foundation for Basic Research grant RFBR-13-04-00700 and the Molecular and Cell Biology Program of the Russian Academy of Sciences (to Y.A.N. and D.V.I.), and the OPTEC research grant (to D.V.I. and L.S.S.).",

year = "2013",

month = dec,

doi = "10.1007/s00441-013-1710-y",

language = "English (US)",

volume = "354",

pages = "783--792",

journal = "Cell and Tissue Research",

issn = "0302-766X",

publisher = "Springer Verlag",

number = "3",

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TY - JOUR

T1 - Arp2/3 complex inhibitors adversely affect actin cytoskeleton remodeling in the cultured murine kidney collecting duct M-1 cells

AU - Ilatovskaya, Daria V.

AU - Chubinskiy-Nadezhdin, Vladislav

AU - Pavlov, Tengis S.

AU - Shuyskiy, Leonid S.

AU - Tomilin, Viktor

AU - Palygin, Oleg

AU - Staruschenko, Alexander

AU - Negulyaev, Yuri A.

N1 - Funding Information: Acknowledgments The authors would like to acknowledge Dr. Grigoriy Stein (Institute of Cytology RAS) for help with microscopy experiments and sincerely thank Vladislav Levchenko (Medical College of Wisconsin) for help with the MTT cytotoxicity assay and critical reading of the manuscript. We also acknowledge the help of Glenn Slocum and Bradley Endres (Medical College of Wisconsin) for helpful discussion and correction of the manuscript. The laboratory of Dr. A.N. Tomilin (Institute of Cytology RAS, St. Petersburg) is recognized for sharing the antibodies for immunofluorescence. This research was supported by R01HL108880 from the National Heart, Lung, and Blood Institute and the American Diabetes Association grant 1-10-BS-168 (to A.S.), Russian Foundation for Basic Research grant RFBR-13-04-00700 and the Molecular and Cell Biology Program of the Russian Academy of Sciences (to Y.A.N. and D.V.I.), and the OPTEC research grant (to D.V.I. and L.S.S.).

PY - 2013/12

Y1 - 2013/12

N2 - Dynamic remodeling of the actin cytoskeleton plays an essential role in cell migration and various signaling processes in living cells. One of the critical factors that controls the nucleation of new actin filaments in eukaryotic cells is the actin-related protein 2/3 (Arp2/3) complex. Recently, two novel classes of small molecules that bind to different sites on the Arp2/3 complex and inhibit its ability to nucleate F-actin have been discovered and described. The current study aims at investigating the effects of CK-0944666 (CK-666) and its analogs (CK-869 and inactive CK-689) on the reorganization of the actin microfilaments in the cortical collecting duct cell line, M-1. We show that treatment with CK-666 and CK869 results in the reorganization of F-actin and drastically affects cell motility rate. The concentrations of the compounds used in this study (100-200 μM) neither cause loss of cell viability nor influence cell shape or monolayer integrity; hence, the effects of described compounds were not due to structural side effects. Therefore, we conclude that the Arp2/3 complex plays an important role in cell motility and F-actin reorganization in M-1 cells. Furthermore, CK-666 and its analogs are useful tools for the investigation of the Arp2/3 complex.

AB - Dynamic remodeling of the actin cytoskeleton plays an essential role in cell migration and various signaling processes in living cells. One of the critical factors that controls the nucleation of new actin filaments in eukaryotic cells is the actin-related protein 2/3 (Arp2/3) complex. Recently, two novel classes of small molecules that bind to different sites on the Arp2/3 complex and inhibit its ability to nucleate F-actin have been discovered and described. The current study aims at investigating the effects of CK-0944666 (CK-666) and its analogs (CK-869 and inactive CK-689) on the reorganization of the actin microfilaments in the cortical collecting duct cell line, M-1. We show that treatment with CK-666 and CK869 results in the reorganization of F-actin and drastically affects cell motility rate. The concentrations of the compounds used in this study (100-200 μM) neither cause loss of cell viability nor influence cell shape or monolayer integrity; hence, the effects of described compounds were not due to structural side effects. Therefore, we conclude that the Arp2/3 complex plays an important role in cell motility and F-actin reorganization in M-1 cells. Furthermore, CK-666 and its analogs are useful tools for the investigation of the Arp2/3 complex.

KW - Actin filaments

KW - Arp2/3 complex

KW - CK-0944666

KW - Cell motility

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JO - Cell and Tissue Research

JF - Cell and Tissue Research

IS - 3

ER -

Arp2/3 complex inhibitors adversely affect actin cytoskeleton remodeling in the cultured murine kidney collecting duct M-1 cells

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