A fluorescent resonant energy transfer–based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows

Chew, Teng Leong; Wolf, Wendy A.; Gallagher, Patricia J.; Matsumura, Fumio; Chisholm, Rex L.

doi:10.1083/jcb.200110161

Cited by 112 publications

(92 citation statements)

References 47 publications

(81 reference statements)

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“…There is strong evidence supporting the importance of ROCK as well as MLCK during cytokinesis (Poperechnaya et al, 2000;Matsumura et al, 2001;Chew et al, 2002;Matsumura, 2005). However, we find that although both signaling mechanisms are required for cytokinesis, they are required at different times.…”

Section: Signaling Pathways Regulating Myosin II Activationmentioning

confidence: 47%

“…Studies of cylindrical sea urchin eggs suggested the presence of a molecular switch that signals the onset of cytokinesis and which is activated at the metaphase-anaphase transition (Shuster and Burgess, 2002b). It is possible that this switch identified in earlier experiments is related to the rise in cortical contractility observed in our compression assays.There is strong evidence supporting the importance of ROCK as well as MLCK during cytokinesis (Poperechnaya et al, 2000;Matsumura et al, 2001;Chew et al, 2002;Matsumura, 2005). However, we find that although both signaling mechanisms are required for cytokinesis, they are required at different times.…”

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confidence: 47%

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A Global, Myosin Light Chain Kinase-dependent Increase in Myosin II Contractility Accompanies the Metaphase–Anaphase Transition in Sea Urchin Eggs

Lucero

Stack

Bresnick

et al. 2006

MBoC

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Myosin II is the force-generating motor for cytokinesis, and although it is accepted that myosin contractility is greatest at the cell equator, the temporal and spatial cues that direct equatorial contractility are not known. Dividing sea urchin eggs were placed under compression to study myosin II-based contractile dynamics, and cells manipulated in this manner underwent an abrupt, global increase in cortical contractility concomitant with the metaphase-anaphase transition, followed by a brief relaxation and the onset of furrowing. Prefurrow cortical contractility both preceded and was independent of astral microtubule elongation, suggesting that the initial activation of myosin II preceded cleavage plane specification. The initial rise in contractility required myosin light chain kinase but not Rho-kinase, but both signaling pathways were required for successful cytokinesis. Last, mobilization of intracellular calcium during metaphase induced a contractile response, suggesting that calcium transients may be partially responsible for the timing of this initial contractile event. Together, these findings suggest that myosin II-based contractility is initiated at the metaphaseanaphase transition by Ca 2؉ -dependent myosin light chain kinase (MLCK) activity and is maintained through cytokinesis by both MLCK-and Rho-dependent signaling. Moreover, the signals that initiate myosin II contractility respond to specific cell cycle transitions independently of the microtubule-dependent cleavage stimulus. INTRODUCTIONAt its most fundamental level, cytokinesis in animal cells is brought about by regional differences in cortical contractility that drives partitioning of the cytoplasm into two daughter cells (Wang, 2005). Force generation is accomplished by the transient assembly of a contractile ring, and recent genetic and proteomic approaches have significantly increased our knowledge of ring components and regulatory molecules (Echard et al., 2004;Eggert et al., 2004;Skop et al., 2004). Studies in yeast using green fluorescent protein-tagged ring components have demonstrated that ring components are recruited to the division site in a hierarchical manner that involves the progressive assembly of more complex structures (Lippincott and Li, 1998;Wu et al., 2003;Wu and Pollard, 2005). In contrast, the temporal sequence in which ring components are recruited to the division site in animal cells is not as well resolved; thus, it remains unclear whether the contractile ring assembles in a similar sequential manner. Moreover, it is not known how ring assembly is coordinated in space and time with chromosome segregation. Thus, although great advances have been made in our understanding of the contractile ring itself, fundamental gaps remain in our understanding of the spatiotemporal regulation of cytokinesis.The mechanical nature of cytokinesis was appreciated by early cell biologists (Wilson, 1928;Rappaport, 1996), and for decades investigators have taken advantage of the large size and regular geometry of echinoderm eggs to st...

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Section: Signaling Pathways Regulating Myosin II Activationmentioning

confidence: 47%

mentioning

confidence: 47%

A Global, Myosin Light Chain Kinase-dependent Increase in Myosin II Contractility Accompanies the Metaphase–Anaphase Transition in Sea Urchin Eggs

Lucero

Stack

Bresnick

et al. 2006

MBoC

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“…Similar approaches have been used to explore the regulated conformations of proteins, such as kinesin and myosin light chain kinase activation (40,41). Before testing our model on ␤-catenin, robust positive and negative control constructs were designed to assess whether predicted spatial relationships could be quantified and confirmed (Fig.…”

Section: Resultsmentioning

confidence: 99%

The Terminal Region of β-Catenin Promotes Stability by Shielding the Armadillo Repeats from the Axin-scaffold Destruction Complex

Chew

Maher

et al. 2009

Journal of Biological Chemistry

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Post-translational stabilization of ␤-catenin is a key step in Wnt signaling, but the features of ␤-catenin required for stabilization are incompletely understood. We show that forms of ␤-catenin lacking the unstructured C-terminal domain (CTD) show faster turnover than full-length or minimally truncated ␤-catenins. Mutants that exhibit faster turnover show enhanced association with axin in co-transfected cells, and excess CTD polypeptide can compete binding of the ␤-catenin armadillo (arm) repeat domain to axin in vitro, indicating that the CTD may restrict ␤-catenin binding to the axin-scaffold complex. Fluorescent resonance energy transmission (FRET) analysis of cyan fluorescent protein (CFP)-arm-CTD-yellow fluorescent protein ␤-catenin reveals that the CTD of ␤-catenin can become spatially close to the N-terminal arm repeat region of ␤-catenin. FRET activity is strongly diminished by the coexpression of ␤-catenin binding partners, indicating that an unliganded groove is absolutely required for an orientation that allows FRET. Amino acids 733-759 are critical for ␤-catenin FRET activity and stability. These data indicate that an N-terminal orientation of the CTD is required for ␤-catenin stabilization and suggest a model where the CTD extends toward the N-terminal arm repeats, shielding these repeats from the ␤-catenin destruction complex.The protein ␤-catenin serves two fundamental roles in the formation and maintenance of tissues. At the cell surface, ␤-catenin binds the cytoplasmic domain of cadherin-type adhesion receptors, allowing cells to engage their neighbors through robust intercellular adhering junctions. In the cytoplasm and nucleus, a cadherin-independent pool of ␤-catenin interacts with TCF 3 -type transcription factors to activate genes that produce cells with distinct identities. The abundance of this cytosolic/nuclear pool of ␤-catenin is largely determined by the presence of extracellular Wnt factors, which initiate a signaling cascade that prevents the continual destruction of cadherin-free ␤-catenin.The destruction of ␤-catenin is carried out by a highly coordinated series of phosphorylation events. In the absence of Wnt, the N terminus of ␤-catenin is sequentially phosphorylated by casein kinase 1␣ and glycogen synthase kinase 3␤ (GSK3␤) (1, 2). Phosphorylation of ␤-catenin by GSK at residues serine 33 and 37 allows recognition by the E3-ligase component ␤TrCP, which when part of the SCF ␤TrCP complex, catalyzes the ubiquitylation and rapid degradation of ␤-catenin (3). This phosphorylation-dependent degradation of ␤-catenin depends on the scaffold protein, axin, and the tumor suppressor APC. Axin has binding sites for casein kinase 1␣, GSK3␤, ␤-catenin, and APC, so that phosphorylation of axin by casein kinase 1␣ and GSK3␤ increases binding to ␤-catenin (4 -6), allowing the N-terminal region of ␤-catenin to be a more efficient substrate of casein kinase 1␣ and GSK3␤ (7). Axin also promotes phosphorylation of APC by casein kinase 1⑀ and GSK3␤ (8,9), increasing the affinity of APC for ␤-ca...

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“…Because RLC kinases including MLCK [29,30], ROK [31,32], and citron kinase [33] are localized in cleavage furrows, and because regulation of RLC kinases could theoretically control RLC phosphorylation, MP regulation may not be essential for cytokinesis. It should be noted, however, that contractile rings are dynamic structures that must contract and, at the same time, disassemble during cytokinesis [34,35].…”

Section: Mypt1 In Cell Divisionmentioning

confidence: 99%

Myosin phosphatase target subunit: Many roles in cell function

Matsumura

Hartshorne

2008

Biochemical and Biophysical Research Communications

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Phosphorylation of myosin II is important in many aspects of cell function and involves a myosin kinase, e.g. myosin light chain kinase, and a myosin phosphatase (MP). MP is regulated by the myosin phosphatase target subunit (MYPT1). The domain structure, properties, and genetic analyses of MYPT1 and its isoforms are outlined. MYPT1 binds the catalytic subunit of type 1 phosphatase, δ isoform, and also acts as an interactive platform for many other proteins. A key reaction for MP is with phosphorylated myosin II and the first process shown to be regulated by MP was contractile activity of smooth muscle. In cell division and cell migration myosin II phosphorylation also plays a critical role and these are discussed. However, based on the wide range of partners for MYPT1 it is likely that MP is implicated with substrates other than myosin II. Open questions are whether the diverse functions of MP reflect different cellular locations and/or specific roles for the MYPT1 isoforms.

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A fluorescent resonant energy transfer–based biosensor reveals transient and regional myosin light chain kinase activation in lamella and cleavage furrows

Cited by 112 publications

References 47 publications

A Global, Myosin Light Chain Kinase-dependent Increase in Myosin II Contractility Accompanies the Metaphase–Anaphase Transition in Sea Urchin Eggs

A Global, Myosin Light Chain Kinase-dependent Increase in Myosin II Contractility Accompanies the Metaphase–Anaphase Transition in Sea Urchin Eggs

The Terminal Region of β-Catenin Promotes Stability by Shielding the Armadillo Repeats from the Axin-scaffold Destruction Complex

Myosin phosphatase target subunit: Many roles in cell function

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