Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration

Joshua C Sandquist; Katherine I Swenson; Kris A Demali; Keith Burridge; Anthony R Means

doi:10.1074/jbc.M605343200

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Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration

Journal article

Open access

Peer reviewed

Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration

Joshua C Sandquist, Katherine I Swenson, Kris A Demali, Keith Burridge and Anthony R Means

The Journal of biological chemistry, Vol.281(47), pp.35873-35883

11/24/2006

DOI: 10.1074/jbc.M605343200

PMID: 17020881

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Abstract

The actin-myosin cytoskeleton is generally accepted to produce the contractile forces necessary for cellular processes such as cell rounding and migration. All vertebrates examined to date are known to express at least two isoforms of non-muscle myosin II, referred to as myosin IIA and myosin IIB. Studies of myosin IIA and IIB in cultured cells and null mice suggest that these isoforms perform distinct functions. However, how each myosin II isoform contributes individually to all the cellular functions attributed to "myosin II" has yet to be fully characterized. Using isoform-specific small-interfering RNAs, we found that depletion of either isoform resulted in opposing migration phenotypes, with myosin IIA- and IIB-depleted cells exhibiting higher and lower wound healing migration rates, respectively. In addition, myosin IIA-depleted cells demonstrated impaired thrombin-induced cell rounding and undertook a more motile morphology, exhibiting decreased amounts of stress fibers and focal adhesions, with concomitant increases in cellular protrusions. Cells depleted of myosin IIB, however, were efficient in thrombin-induced cell rounding, displayed a more retractile phenotype, and maintained focal adhesions but only in the periphery. Last, we present evidence that Rho kinase preferentially regulates phosphorylation of the regulatory light chain associated with myosin IIA. Our data suggest that the myosin IIA and IIB isoforms are regulated by different signaling pathways to perform distinct cellular activities and that myosin IIA is preferentially required for Rho-mediated contractile functions.

Phosphorylation

Signal Transduction

Protein-Serine-Threonine Kinases - physiology

Humans

Intracellular Signaling Peptides and Proteins - metabolism

Microscopy, Video

Wound Healing

RNA - chemistry

Muscle Contraction

Thrombin - chemistry

Protein Isoforms

rho-Associated Kinases

Cell Line, Tumor

Intracellular Signaling Peptides and Proteins - physiology

Myosin Type II - chemistry

Microscopy, Fluorescence

Protein-Serine-Threonine Kinases - metabolism

Cell Movement

RNA, Small Interfering - metabolism

Details

Title: Subtitle: Rho kinase differentially regulates phosphorylation of nonmuscle myosin II isoforms A and B during cell rounding and migration
Creators: Joshua C Sandquist - Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina 27710, USA
Katherine I Swenson
Kris A Demali
Keith Burridge
Anthony R Means
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.281(47), pp.35873-35883
DOI: 10.1074/jbc.M605343200
PMID: 17020881
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: CA 082845 / NCI NIH HHS GM 29860 / NIGMS NIH HHS
Language: English
Date published: 11/24/2006
Academic Unit: Dermatology; Fraternal Order of Eagles Diabetes Research Center; Biochemistry and Molecular Biology
Record Identifier: 9984024404102771

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