G1/S control of anchorage-independent growth in the fibroblast cell cycle.

Guadagno, Thomas M.; Assoian, Richard K.

doi:10.1083/jcb.115.5.1419

Cited by 117 publications

(84 citation statements)

References 52 publications

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“…As we noted previously (Guadagno and Assoian, 1991), the shared features include: (a) blocked DNA synthesis despite normal cell cycle progression through most of G1, (b) selective failure to express G1/S cell cycle genes, (c) continued increases in cell size despite lack of cell cycle progression, and (d) arrest before activation of the cdc2-1ike kinase.…”

Section: Ell Cycle Progression In Nontransformed Fibroblastsmentioning

confidence: 93%

“…As discussed above, our previous studies on cell cycle control of anchorage-independent growth (Guadagno and Assoian, 1991) showed that nonadherent, G0-synchronized fibroblasts arrest in late G1 with a phenotype that resembles arrest at START in S. cerevisiae. To determine if there are additional attachment-dependent cell cycle transitions, we prepared parallel cultures of adherent and nonadherent NRK and NIH-3T3 fibroblasts that had been synchronized at the G1/S interface by exposure to hydroxyurea (Draetta and Beach, 1988;Pledger et al, 1977;DeCaprio et al, 1989).…”

Section: Hydroxyurea Synchronization Establishes Gi/s As the Sole Attmentioning

confidence: 99%

“…We have recently extended these studies on attachment control of the fibroblast cell cycle and characterized a cell cycle block to anchorage-independent growth in detail (Guadagno and Assoian, 1991). In contrast to studies with standard soft agar cultures, our experiments are based on use of a preparative suspension culture system that allows for easy recovery and analysis of cells.…”

Section: Ell Cycle Progression In Nontransformed Fibroblastsmentioning

confidence: 99%

“…After removal of the drug, S-and G2/M-phase progression was monitored by [3H]thymidine incorporation and changes in cell number, respectively. Results from these studies ( closed circles) 35-mm dishes as described (Guadagno and Assoian, 1991).…”

Section: Hydroxyurea Synchronization Establishes Gi/s As the Sole Attmentioning

confidence: 99%

“…When applicable, mitogen-treated adherent cells, or TGF-~ treated nonadherent cells, were cultured in parallel as positive control. See Guadagno and Assoian (1991) for methodologic details.…”

Section: Characterization Of Anchorage-independent Growth In Preparatmentioning

confidence: 99%

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A cell cycle and mutational analysis of anchorage-independent growth: cell adhesion and TGF-beta 1 control G1/S transit specifically

Han

Guadagno

Dalton

et al. 1993

The Journal of Cell Biology

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Abstract. We have examined cell cycle control of anchorage-independent growth in nontransformed fibroblasts. In previous studies using Go-synchronized NRK and NIH-3T3 cells, we showed that anchorageindependent growth is regulated by an attachmentdependent transition at G1/S that resembles the START control point in the cell cycle of Saccharomyces cerevisiae. In the studies reported here, we have synchronized NRK and NIH-3T3 fibroblasts immediately after this attachment-dependent transition to determine if other portions of the fibroblast cell cycle are similarly regulated by adhesion. Our results show that S-, G2-, and M-phase progression proceed in the absence of attachment. Thus, we conclude that the adhesion requirement for proliferation of these cells can be explained in terms of the single START-like transition. In related studies, we show that TGF-/~I overrides the attachment-dependent transition in NRK and AKR-2B fibroblasts (lines in which TGF-/$1 induces anchorageindependent growth), but not in NIH-3T3 or Balb/c 3T3 fibroblasts (lines in which TGF-/31 fails to induce anchorage-independent growth). These results show that (a) adhesion and TGF-/31 can have similar effects in stimulating cell cycle progression from G1 to S and (b) the differential effects of TGF-fll on anchorageindependent growth of various fibroblast lines are directly reflected in the differential effects of the growth factor at G1/S. Finally, we have randomly mutagenized NRK fibroblasts to generate mutant lines that have lost their attachment/TGF-~l requirement for G1/S transit while retaining their normal mitogen requirements for proliferation. These clones, which readily proliferate in mitogen-supplemented soft agar, appear non-transformed in monolayer: they are well spread, nonrefractile, and contact inhibited. The existence of this new fibroblast phenotype demonstrates (a) that the growth factor and adhesion/TGF-~/1 requirements for cell cycle progression are genetically separable, (b) that the two major control points in the fibroblast cell cycle (G0/G1 and G1/S) are regulated by distinct extracellular signals, and (c) that the genes regulating anchorage-independent growth need not be involved in regulating contact inhibition, focus formation, or growth factor dependence.

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Section: Ell Cycle Progression In Nontransformed Fibroblastsmentioning

confidence: 93%

Section: Hydroxyurea Synchronization Establishes Gi/s As the Sole Attmentioning

confidence: 99%

Section: Ell Cycle Progression In Nontransformed Fibroblastsmentioning

confidence: 99%

Section: Hydroxyurea Synchronization Establishes Gi/s As the Sole Attmentioning

confidence: 99%

Section: Characterization Of Anchorage-independent Growth In Preparatmentioning

confidence: 99%

See 3 more Smart Citations

A cell cycle and mutational analysis of anchorage-independent growth: cell adhesion and TGF-beta 1 control G1/S transit specifically

Han

Guadagno

Dalton

et al. 1993

The Journal of Cell Biology

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Cytoarchitecture and cell growth control

Slack¹,

Higgins²

1996

Cell Motil. Cytoskeleton

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Attenuation of plasminogen activator inhibitor type-1 promoter activity in serum-stimulated renal epithelial cells by a distal 5? flanking region

Slack

Higgins

1999

Cell Motil. Cytoskeleton

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Urokinase plasminogen activator (u‐PA) and its fast acting type‐1 inhibitor (PAI‐1) localize to cellular focal adhesive structures and the ajoining proximal undersurface region, respectively (Kutz et al., J. Cell. Physiol. 176:8–18, 1997). PAI‐1 may function in this locale to modulate pericellular proteolytic activity, cell‐to‐substrate adhesion, or matrix‐dependent motility. While PAI‐1 synthesis is regulated in an immediate‐early response manner in growth “activated” renal cells coincident with cytoskeletal restructuring, adhesive influences both repress the amplitude and prolong the time course of serum‐induced PAI‐1 transcription (Ryan et al., Biochem. J. 314:1041–1046, 1996). To identify potential adhesion‐responsive elements within the PAI‐1 gene that function in this complex mode of expression control, reporter constructs containing defined directionally deleted PAI‐1 5′ genomic fragments cloned upstream of a CAT gene were employed in transient transfection assays. A 483‐bp distal PAI‐1 flanking segment (corresponding to nucleotides −2395 to −1912) conferred significant adhesion‐dependent attenuation on serum‐induced PAI‐1 transcription. This 483‐bp distal PAI‐1 segment functioned as a repressor of reporter (CAT) activity under both adhesive and suspension culture conditions, however, when ligated upstream of either an SV40 promoter/enhancer or a minimal PAI‐1 promoter. These data suggest that repressor elements located between −2395 and −1912 bp interact with more proximal adhesion‐dependent regulatory elements to affect PAI‐1 expression attenuation during serum stimulation of adherent renal epithelial cells. Cell Motil. Cytoskeleton 44:168–176, 1999. © 1999 Wiley‐Liss, Inc.

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G1/S control of anchorage-independent growth in the fibroblast cell cycle.

Cited by 117 publications

References 52 publications

A cell cycle and mutational analysis of anchorage-independent growth: cell adhesion and TGF-beta 1 control G1/S transit specifically

A cell cycle and mutational analysis of anchorage-independent growth: cell adhesion and TGF-beta 1 control G1/S transit specifically

Cytoarchitecture and cell growth control

Attenuation of plasminogen activator inhibitor type-1 promoter activity in serum-stimulated renal epithelial cells by a distal 5? flanking region

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