Transformation parameters and pp60src localization in cells infected with partial transformation mutants of Rous sarcoma virus.

Rohrschneider, Larry R.; Rosok, M J

doi:10.1128/mcb.3.4.731

Cited by 69 publications

(57 citation statements)

References 34 publications

(61 reference statements)

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“…All these proteins are localized at focal contacts where they form complexes with integrins and other proteins to anchor the actin fibers to the site of cell contact (9,10). In v-src-transformed cells, pp60 v-src is localized at the adhesion plaques, and the amount of actin stress fibers is greatly reduced (11). These findings indicate that regulation of Src family kinases is essential to integrity of the actin cytoskeleton (1).…”

mentioning

confidence: 68%

Activation of pp60 Depending on Cell Density in PC12h Cells

Kobayashi¹,

Okumura²,

Nakamoto³

et al. 1997

Journal of Biological Chemistry

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The Src family tyrosine kinases and their substrates are involved in cell-cell and cell-matrix interactions. We found that in PC12h cells, an increase of cell density enhanced the tyrosine phosphorylation levels of several intracellular proteins including p130 cas . Because it is a possible substrate for Src family kinases, we measured pp60 c-src activity and found that it was higher in high density cultures than in low density cultures. This phenomenon was also observed in PC12 (the parental cell line of the PC12h subclone), Balb/c 3T3, Swiss 3T3, and Hela cells. One of the possible mechanisms regulating the kinase activity of pp60 c-src is the phosphorylation and dephosphorylation of its negative regulatory site located at its C terminus. However, the tyrosine phosphorylation level of the regulatory site did not change depending on cell density. Subcellular fractionation showed that in high density culture, pp60 c-src was translocated from detergent-soluble to detergent-insoluble fractions. These results suggest that cell-cell interaction might induce the activation of pp60 c-src without changing its tyrosine phosphorylation levels.

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mentioning

confidence: 68%

Activation of pp60 Depending on Cell Density in PC12h Cells

Kobayashi¹,

Okumura²,

Nakamoto³

et al. 1997

Journal of Biological Chemistry

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“…The subsequent disruption of stress fibers suggests that vinculin and adhesion plaques may play a crucial role in their stability. On the other hand, the extent of vinculin phosphorylation seems limited in transformed cells (31), and increases in vinculin phosphorylation alone appears insufficient to cause dissolution of stress fibers (28).…”

Section: Discussionmentioning

confidence: 99%

Reorganization of alpha-actinin and vinculin induced by a phorbol ester in living cells.

Meigs¹,

Wang²

1986

The Journal of Cell Biology

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Abstract. We have used fluorescent analogue cytochemistry, image intensification, and digital image processing to examine the redistribution of alphaactinin and vinculin in living cultured African green monkey kidney (BSC-1) cells treated with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA). Before treatment, microinjected alpha-actinin shows characteristic distribution along stress fibers and at adhesion plaques; vinculin is localized predominantly at adhesion plaques. Soon after the addition of TPA, highly dynamic membrane ruffles begin to form. These incorporate a large amount of alpha-actinin but little vinculin. Alpha-actinin is subsequently depleted, more or less uniformly, from stress fibers. Disrupted stress fibers often fragment into aggregates and move into the perinuclear region. Careful analyses of fluorescence intensity distribution indicate that alphaactinin is depleted more rapidly from adhesion plaques than from stress fibers. Furthermore, the depletion of alpha-actinin from adhesion plaques is also faster than either the depletion of vinculin or the disappearance of focal contacts. These observations indicate that TPA may initiate disruption of stress fibers by interfering with a link between alpha-actinin and vinculin, causing alpha-actinin to be preferentially depleted from adhesion plaques. MANY cultured nonmuscle cells contain a well-organized set of stress fibers, which are large bundles of actin filaments associated with various accessory proteins such as alpha-actinin, myosin, and tropomyosin (for a review see reference 22). One of the most consistent characteristics of cells infected with transforming viruses, cells treated with tumor-promoting phorbol esters, and cells stimulated with growth factors, is the disorganization of stress fibers (for a review see reference 33). Since all of these agents stimulate various kinds of protein kinases (6,8,9,12,17,25), it is possible that a common pathway is followed during the disruption process.The reorganization of actin and vinculin induced by the tumor-promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA),~ has recently been studied using fluorescent phalloidin staining, immunofluorescence, and electron microscopy. Shortly after treatment, large actin-containing rufties appear and stress fibers start to disappear from the cytoplasm (26, 30). Vinculin can be detected at the ends of stress fibers, where adhesion plaques are located, during the early stage of disruption. At steady state, the cell contains prominent ruffles and numerous aggregates of actin and vinculin. Although these studies have shown the dramatic effects of Abbreviations used in this paper." DMSO, dimethyl sulfoxide; FITC, fluorescein isothiocyanate; IATR, iodoacetamidotetramethylrhodamine; IRM, interference reflection microscopy; 4-alpha-PDD, 4-alpha-phorbol 12,13-didecanoate: TPA, 12-O-tetradecanoylphorbol-13-acetate.TPA, the actual process of structural reorganization can only be speculated upon since most observations were performed on different, ...

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“…It is then likely that 'adhesion plaques' and the interacting cytoskeletal structures are involved in the transformation process, as supported by the fact that pp6Osrc binds here selectively (Rohrschneider and Rosok, 1983). pp6Osrc may exert its kinase activity locally on substrates, other than vinculin, corresponding to some or all the above described cytoskeleton-associated molecules.…”

Section: Phosphorylation In Vitromentioning

confidence: 98%

“…This is in agreement with data showing that the tyrosine-phosphorylated form of pp60src is membrane bound, while the serine-phosphorylated form is found in the detergent-soluble fraction (Courtneidge and Bishop, 1982 , and was found in the detergent-treated cell fractions used in this work for the kinase reaction. Since a limited amount of vinculin has been demonstrated to be phosphorylated at tyrosine residues in vivo in RSV-transformed cells (Sefton et al, 1981) and this molecule and pp6Osrc co-exist within defined structures of the ventral plasma membrane, called 'adhesion plaques' Rohrschneider and Rosok, 1983), the issue of whether vinculin is a significant target for pp6Osrc has been debated (Hynes, 1982). If this were the case, the conditions used here for the kinase reaction would allow quantitative phosphorylation of the vinculin associated with the detergent-insoluble cell fraction.…”

Section: Phosphorylation In Vitromentioning

confidence: 99%

Detection of phosphotyrosine-containing proteins in the detergent-insoluble fraction of RSV-transformed fibroblasts by azobenzene phosphonate antibodies.

Comoglio

Renzo

Tarone³

et al. 1984

The EMBO Journal

149

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conditions allowing the kinase reaction catalyzed by pp6Osrc. Phosphoproteins of closely related mol. wts. were immunoprecipitated from RSV-transformed avian fibroblasts. The radioactivity co-migrated with authentic phosphotyrosine in two-dimensional chromatography. The 60-kd protein comigrated with pp6Osrc, while the identity between the 130-kd protein and vinculin was disproved by the lack of crossreaction with appropriate antisera. In tiansformed mouse and duck fibroblasts ABP antibodies, employed in indirect immunofluorescence microscopy, stained diffusely the cytoplasm and intensely decorated restricted areas of the ventral cell plasma membrane. These data show that antibodies reacting with phosphotyrosine may be usefully employed in the identification and in the intracellular localization of molecules that are potential targets of the pp6Osrc protein kinase.

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Transformation parameters and pp60src localization in cells infected with partial transformation mutants of Rous sarcoma virus.

Cited by 69 publications

References 34 publications

Activation of pp60 Depending on Cell Density in PC12h Cells

Activation of pp60 Depending on Cell Density in PC12h Cells

Reorganization of alpha-actinin and vinculin induced by a phorbol ester in living cells.

Detection of phosphotyrosine-containing proteins in the detergent-insoluble fraction of RSV-transformed fibroblasts by azobenzene phosphonate antibodies.

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