Actin and serum response factor transduce physical cues from the microenvironment to regulate epidermal stem cell fate decisions

Connelly, John T.; Gautrot, Julien E.; Trappmann, Britta; Tan, David Wei-Min; Donati, Giacomo; Huck, Wilhelm T. S.; Watt, Fiona M.

doi:10.1038/ncb2074

Cited by 418 publications

(455 citation statements)

References 38 publications

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“…Other mechanical cues correlated with fate commitment of mesenchymal stem cells are cell shape and the mean stress in the cytoskeleton (15). Because actin transfers physical cues between the cell exterior and interior, it likely plays a major role in stem cell linage commitment (14,25). However, there have been no measurements available on the forces in specific structural proteins in adult cell reprogramming and dedifferentiation.…”

Section: Resultsmentioning

confidence: 99%

“…Here we present, to our knowledge, the first probe to measure force in a polymeric protein, f-actin, that fulfills many structural and motor functions in cells. For example, f-actin forms filaments that bridge chromatin domains in the nucleus to the cell membrane and the extracellular matrix, providing a path for the well-known effects of mechanical stress on gene expression (13), stem cell differentiation, and mobility (14,15).…”

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confidence: 99%

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Actin stress in cell reprogramming

Guo

Wang

Sachs

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

103

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Cell mechanics plays a role in stem cell reprogramming and differentiation. To understand this process better, we created a genetically encoded optical probe, named actin-cpstFRET-actin (AcpA), to report forces in actin in living cells in real time. We showed that stemness was associated with increased force in actin. We reprogrammed HEK-293 cells into stem-like cells using no transcription factors but simply by softening the substrate. However, Madin-Darby canine kidney (MDCK) cell reprogramming required, in addition to a soft substrate, Harvey rat sarcoma viral oncogene homolog expression. Replating the stem-like cells on glass led to redifferentiation and reduced force in actin. The actin force probe was a FRET sensor, called cpstFRET (circularly permuted stretch sensitive FRET), flanked by g-actin subunits. The labeled actin expressed efficiently in HEK, MDCK, 3T3, and bovine aortic endothelial cells and in multiple stable cell lines created from those cells. The viability of the cell lines demonstrated that labeled actin did not significantly affect cell physiology. The labeled actin distribution was similar to that observed with GFPtagged actin. We also examined the stress in the actin cross-linker actinin. Actinin force was not always correlated with actin force, emphasizing the need for addressing protein specificity when discussing forces. Because actin is a primary structural protein in animal cells, understanding its force distribution is central to understanding animal cell physiology and the many linked reactions such as stress-induced gene expression. This new probe permits measuring actin forces in a wide range of experiments on preparations ranging from isolated proteins to transgenic animals.actin | force probe | stem cell | reprogramming | cell mechanics C ells have only three sources of free energy-chemical, electrical, and mechanical potential (1)-and life is driven by the flow of energy between them. The flow of mechanical energy has not been well studied in living cells outside of muscle, yet all cells are subject to endogenous and exogenous mechanical forces. The lack of progress in measuring these forces (stress) has primarily been due to a lack of suitable probes, but that has now been remedied with the development of genetically coded FRETbased force sensors (2). The literature shows many macroscopic effects of mechanical stress on cellular processes including cell motility, embryogenesis, stem cell replication and differentiation (3, 4), bone and muscle homeostasis, gene expression, protein folding (5), and membrane potential (6). However, these studies lacked the ability to measure stress in specific structural proteins, and the analyses have tended to treat cytoskeletal stresses as uniform, which we now know is not true (2, 7). We, and other groups (8), have shown that the distribution of forces is nonuniform in both time and space and protein specificity (9).The force sensors consist of a FRET pair (typically CFP/ YFP) connected by a protein linker, a biological analog of a mechanic...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Actin stress in cell reprogramming

Guo

Wang

Sachs

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

103

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“…FHOD3 likely activated SRF via actin dynamics, because the response was effectively blocked by latrunculin A and latrunculin B (Figure 2A), both of which are known to inhibit actin polymerization and thus increase the level of Gactin, leading to impairment of SRF activity. 17, 21 In contrast to FHOD3-WT, a mutant mouse Fhod3 construct carrying the I1127A mutation, which has been demonstrated to be defective in in-vivo actin-assembling activity, 3 failed to induce the activation of SRF-dependent transcription ( Figure 2B). Therefore, FHOD3 appeared to activate SRF in a manner dependent on actin-assembling activity.…”

Section: Identification Of Fhod3 Mutations In Dcmmentioning

confidence: 99%

Dilated Cardiomyopathy-Associated <i>FHOD3</i> Variant Impairs the Ability to Induce Activation of Transcription Factor Serum Response Factor

Arimura

Takeya

Ishikawa

et al. 2013

Circ J

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2990ARIMURA T et al. Circulation JournalOfficial Journal of the Japanese Circulation Society http://www. j-circ.or.jp ilated cardiomyopathy (DCM) is a primary cardiac disorder caused by functional abnormalities in cardiomyocytes and is characterized by ventricular chamber dilation with decreased contractility. DCM is a major cause of chronic heart failure and the most common indication for cardiac transplantation. Various etiologies include genetic abnormalities, viral infections, alcohol, mitochondrial dysfunction and metabolic disorders. 1,2 Most of the genetic causes are mutations in genes for sarcomeric proteins, including contractile elements, sarcolemma elements, Z-disc elements, and Z-I region components, which play key roles in the generation and transmission of contractile force. 2 Editorial p 2879Formin homology 2 domain containing 3 (FHOD3) is a member of the formin family. We have previously reported that FHOD3 is a sarcomeric protein that is predominantly expressed in the heart and plays an essential role in the regulation of actin assembly and sarcomeric organization during myofibrillogenesis. 3 FHOD3 contains multiple domains, including GTPase-binding and diaphanous inhibitory domains at the Nterminus, formin homologous 1 (FH1), formin homologous 2 (FH2) and diaphanous autoregulation (DA) domains at the Cterminus. 4 The FH2 domain mediates actin filament nucleation and polymerization, which are accelerated by FH1-mediated Background: Dilated cardiomyopathy (DCM) is characterized by a dilated left ventricular cavity with systolic dysfunction manifested by heart failure. It has been revealed that mutations in genes for cytoskeleton or sarcomere proteins cause DCM. However, the disease-causing mutations can be found only in far less than half of patients with a family history, indicating that there should be other disease genes for DCM. Formin homology 2 domain containing 3 (FHOD3) is a sarcomeric protein expressed in the heart that plays an essential role in sarcomere organization during myofibrillogenesis. The purpose of this study was to explore a possible novel disease gene for DCM.

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“…3 While TCFs regulate expression of a number of immediate early genes necessary for cell growth and proliferation, 4,5 MRTFs couple SRF-dependent transcription to signals from Rho family GTPases and intracellular actin dynamics. 2,6 MRTFs play an important role in a large number of developmental and physiological processes, including cardiovascular development, 7,8 epithelial differentiation, 9,10 neuronal plasticity [11][12][13] and cell migration. 14,15 In addition, the closely related SRF coactivator myocardin is a candidate tumor suppressor, 16,17 while MRTFs have been implicated in experimental metastasis.…”

Section: Introductionmentioning

confidence: 99%

Myocardin related transcription factors are required for coordinated cell cycle progression

et al. 2013

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Myocardin related transcription factors A and B (MRTFs) activate serum response factor-driven transcription in response to Rho signaling and changes in actin dynamics. Myocardin and MRTFs have been implicated in anti-proliferative effects on a range of cell types. the precise mechanisms, however, remained elusive. We employed double knockdown of MRTF-A and MRTF-B in NIH 3T3 fibroblasts to evaluate its effects on cell cycle progression and proliferation. We show that transient depletion of MRTFs conveys a modest anti-proliferative effect and impinges on normal cell cycle progression, resulting in significantly shortened G 1 phase and slightly extended S and G 2 phase under normal growth conditions. Under serum-starved conditions we observed aberrant entry into the S and G 2 phases without subsequent cell division. this was accompanied by downregulation of cyclin-CDK inhibitors p27Kip1, p18Ink4c and 19Ink4d as well as upregulation of p21Waf1 and cyclin D1. extended knockdown led to increased formation of micronuclei, while cells stably depleted of MRTFs tend to become aneuploid and polyploid. thus, MRTFs are required for accurate cell cycle progression and maintenance of genomic stability in fibroblast cells

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Actin and serum response factor transduce physical cues from the microenvironment to regulate epidermal stem cell fate decisions

Cited by 418 publications

References 38 publications

Actin stress in cell reprogramming

Actin stress in cell reprogramming

Dilated Cardiomyopathy-Associated <i>FHOD3</i> Variant Impairs the Ability to Induce Activation of Transcription Factor Serum Response Factor

Myocardin related transcription factors are required for coordinated cell cycle progression

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