ADP-ribosylation of the G Protein Rho Inhibits Integrin Regulation of Tumor Cell Growth

Udagawa, Taturo; McIntyre, Bradley W.

doi:10.1074/jbc.271.21.12542

Cited by 35 publications

(28 citation statements)

References 49 publications

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“…There have also been many reports suggesting that occupation of the extracellular matrix receptor, integrin, is required for terminal muscle differentiation (21,45,70,74). A recent report has suggested that Rho family proteins work as mediators of integrin signaling (86). Taking these observations and our results together, it is tempting for us to speculate that integrin regulates muscle differentiation via the Rho family proteins.…”

Section: Discussioncontrasting

confidence: 47%

The Rho Family G Proteins Play a Critical Role in Muscle Differentiation

Takano

Komuro

Oka

et al. 1998

Molecular and Cellular Biology

139

127

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The Rho family GTP-binding proteins play a critical role in a variety of cytoskeleton-dependent cell functions. In this study, we examined the role of Rho family G proteins in muscle differentiation. Dominant negative forms of Rho family proteins and RhoGDI, a GDP dissociation inhibitor, suppressed transcription of muscle-specific genes, while mutationally activated forms of Rho family proteins strongly activated their transcription. C2C12 cells overexpressing RhoGDI (C2C12RhoGDI cells) did not differentiate into myotubes, and expression levels of myogenin, MRF4, and contractile protein genes but not MyoD and myf5 genes were markedly reduced in C2C12RhoGDI cells. The promoter activity of the myogenin gene was suppressed by dominant negative mutants of Rho family proteins and was reduced in C2C12RhoGDI cells. Expression of myocyte enhancer binding factor 2 (MEF2), which has been reported to be required for the expression of the myogenin gene, was reduced at the mRNA and protein levels in C2C12RhoGDI cells. These results suggest that the Rho family proteins play a critical role in muscle differentiation, possibly by regulating the expression of the myogenin and MEF2 genes.Myogenic basic helix-loop-helix (bHLH) proteins are master regulatory proteins that activate the transcription of many muscle-specific genes during myogenesis (reviewed in references 60 and 89). Each of the four myogenic bHLH proteins, MyoD (17), myogenin (18, 90), myf5 (7), and MRF4 (8, 47, 67), can activate the skeletal myogenic program when introduced into a variety of cells derived from all three germ layers of the embryo. The bHLH motif mediates dimerization of myogenic factors with ubiquitous bHLH proteins such as E12/E47, and these heterodimeric complexes bind to a conserved DNA sequence known as the E box, which is present in the promoters and enhancers of most muscle-specific genes (54). Myocyte enhancer binding factor 2 (MEF2), which is a member of the MADS box family, also plays an important role in muscle differentiation (reviewed in reference 61). MEF2 activates transcription by binding to the consensus sequence, called the MEF2-binding site, which is also found in the control regions of numerous muscle-specific genes (23, 64). In embryos with loss-of-function mutations of the single mef2 gene in Drosophila (D-mef2), somatic, cardiac, and visceral muscle cells did not differentiate (6,38,66). These results indicated that MEF2 is necessary for the differentiation of all types of muscle cells. MEF2 and myogenic bHLH proteins have been suggested to activate mutual expression in an autoregulatory network and maintain the expression of muscle-specific genes (5,16,31,43,56). Moreover, a recent study demonstrated that MEF2 and myogenic bHLH proteins synergistically activate expressions of muscle-specific genes via protein-protein interactions between DNA-binding domains of these heterologous classes of transcription factors (51). In addition, it has been reported that a variety of factors, such as fibroblast growth factor, transforming grow...

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Section: Discussioncontrasting

confidence: 47%

The Rho Family G Proteins Play a Critical Role in Muscle Differentiation

Takano

Komuro

Oka

et al. 1998

Molecular and Cellular Biology

139

127

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“…9A). We treated cells with the C3 exoenzyme from Clostridium botulinum, which inhibits the function of RhoA, RhoB and RhoC; the other Rho family proteins Rac-1 and Cdc42 are poor C3 substrates (Udagawa and McIntyre, 1996). We also used Y-27632, a specific inhibitor of the Rho-associated coiled-coilforming protein serine/threonine kinase (ROCK) family of protein kinases (Narumiya et al, 2000).…”

Section: Cd8/mcsp Perturbs Intercellular Adhesion and The Actin Cytosmentioning

confidence: 99%

“…Keratinocytes expressing CD8 or CD8/MCSP were incubated with the following pharmacological inhibitors for 6 hours at 37°C: 5 µM Y-27632 (Calbiochem) (Narumiya et al, 2000); 5 µg ml -1 C3 exoenzyme from Clostridium botulinum (Calbiochem) (Udagawa and McIntyre, 1996); and 50 µM LY294002 (Sigma-Aldrich) (Pullen and Thomas, 1997).…”

Section: Function-blocking Experiments With Antibodies and Pharmacolomentioning

confidence: 99%

Role of melanoma chondroitin sulphate proteoglycan in patterning stem cells in human interfollicular epidermis

et al. 2003

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Human interfollicular epidermis is renewed by stem cells that are clustered in the basal layer in a patterned, non-random distribution. Stem cells can be distinguished from other keratinocytes by high expression of β1 integrins and lack of expression of terminal differentiation markers; they divide infrequently in vivo but form actively growing colonies in culture. In a search for additional stem cell markers, we observed heterogeneous epidermal expression of melanoma chondroitin sulphate proteoglycan (MCSP). MCSP was expressed by those keratinocytes with the highest β1 integrin levels. In interfollicular epidermis, expression was confined to non-cycling cells and,in culture, to self-renewing clones. However, fluorescence-activated cell sorting on the basis of MCSP and β1 integrin expression gave no more enrichment for clonogenic keratinocytes than sorting for β1 integrins alone. To interfere with endogenous MCSP, we retrovirally infected keratinocytes with a chimera of the CD8 extracellular domain and the MCSP cytoplasmic domain. CD8/MCSP did not affect keratinocyte proliferation or differentiation but the cohesiveness of keratinocytes in isolated clones or reconstituted epidermal sheets was greatly reduced. CD8/MCSP caused stem cell progeny to scatter without differentiating. CD8/MCSP did not alter keratinocyte motility but disturbed cadherin-mediated cell-cell adhesion and the cortical actin cytoskeleton, effects that could be mimicked by inhibiting Rho. We conclude that MCSP is a novel marker for epidermal stem cells that contributes to their patterned distribution by promoting stem cell clustering.

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“…Therefore, we examined whether Rho is involved in MEF cell migration. Clostridium botulinum C3 exoenzyme is a specific inhibitor of Rho that works by irreversible ADP-ribosylation of Asn-41 in its effector region (19). We previously used this C3 toxin to block serum-induced Rho activation and actin stress fiber formation in MEF cells (11).…”

Section: Deficiency Of Csk Does Not Affect Rac Activation and Lamellimentioning

confidence: 99%

Role of Tyrosine Kinase Csk in G Protein-coupled Receptor- and Receptor Tyrosine Kinase-induced Fibroblast Cell Migration

McGarrigle

Shan

Yang

et al. 2006

Journal of Biological Chemistry

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Tyrosine kinase Csk is essential for mouse embryonic development. Csk knock-out mice died at early stages of embryogenesis (around embryonic day 10). The molecular mechanism for this defect is not completely understood. Here we report that Csk deficiency in mouse embryonic fibroblast cells blocked cell migration induced by lysophosphatidic acid through G protein-coupled receptors, by platelet-derived growth factor and epidermal growth factor through receptor tyrosine kinases, and by serum. Re-expression of Csk in these Csk-deficient cells rescued the migratory phenotype. Furthermore, deletion of Csk did not interfere with Rac activation and lamellipodia formation, but impaired the focal adhesions. Our data demonstrate a critical role for Csk in cell migration.Protein-tyrosine kinase Csk (C-terminal Src kinase) was originally purified as a kinase capable of phosphorylating Src and other Src family kinases at their C-terminal tyrosine residues (1, 2). This phosphorylation suppresses the kinase activity of Src-family tyrosine kinases (such as c-Src, Fyn, and Lyn). Therefore, most physiological studies regarding Csk function have been focused on Csk as a negative regulator of Srcfamily tyrosine kinases. Csk is ubiquitously expressed in mammalian cells and is evolutionarily conserved from the early diverged metazoan Hydra to humans (3). The Csk Ϫ/Ϫ mouse embryos exhibited defects in neurulation, an inability to complete the turning process, and a failure of the allantois to connect with the chorion, preventing the formation of the umbilical cord and placenta (4, 5). The Csk-deficient mouse embryos died around embryonic day 10. Elegant mouse genetic studies revealed that Src Ϫ/Ϫ Csk Ϫ/Ϫ mouse embryos showed partial rescue of Csk Ϫ/Ϫ phenotypes (6). However, Src Ϫ/Ϫ Csk Ϫ/Ϫ mouse embryos still died around embryonic days 10 -11, implying a Src-dependent and -independent function for Csk (6). Indeed, cellular and biochemical studies suggested that Csk also interacts with other proteins, such as proteintyrosine phosphatase (PTP) 2 -PEST (7). Because of the embryonic lethal phenotype of Csk-knock-out mice, the physiological functions of Csk need to be investigated in individual cellular signaling processes and in different tissues and organs.Cell migration underlies many physiological and pathological processes such as embryonic development, wound healing, angiogenesis, inflammation, and neuronal outgrowth (8). Aberrant migration contributes to pathologies such as atherosclerosis, tumor invasion, and metastasis. Cell migration is a dynamic, cyclical process in which a cell extends a protrusion at its front, which in turn attaches to the substratum on which the cell is migrating (9). This is followed by a contraction that moves the cell body forward toward the protrusion, and finally the attachments at the cell rear release as the cell continues to move forward. The cycle is initiated by external signals, which are sensed and communicated to the interior of the cell by specialized receptor proteins in the cell membrane. Foc...

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ADP-ribosylation of the G Protein Rho Inhibits Integrin Regulation of Tumor Cell Growth

Cited by 35 publications

References 49 publications

The Rho Family G Proteins Play a Critical Role in Muscle Differentiation

The Rho Family G Proteins Play a Critical Role in Muscle Differentiation

Role of melanoma chondroitin sulphate proteoglycan in patterning stem cells in human interfollicular epidermis

Role of Tyrosine Kinase Csk in G Protein-coupled Receptor- and Receptor Tyrosine Kinase-induced Fibroblast Cell Migration

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