A Discrete Cell Cycle Checkpoint in Late G1 That Is Cytoskeleton-Dependent and MAP Kinase (Erk)-Independent

doi:10.1006/excr.2002.5504

Experimental Cell Research

Volume 275, Issue 2, 1 May 2002, Pages 255-264

https://doi.org/10.1006/excr.2002.5504 Get rights and content

Abstract

Cell spreading on extracellular matrix and associated changes in the actin cytoskeleton (CSK) are necessary for progression through G₁ and entry into S phase of the cell cycle. Pharmacological disruption of CSK integrity inhibits early mitogenic signaling to the extracellular signal-regulated kinase (Erk) subfamily of the mitogen-activated protein kinases (MAPKs) and arrests the cell cycle in G₁. Here we show that this block of G₁ progression is not simply a consequence of inhibition of the MAPK/Erk pathway but instead it reveals the existence of a discrete CSK-sensitive checkpoint. Use of PD98059 to inhibit MAPK/Erk and cytochalasin D (Cyto D) to disrupt the actin CSK at progressive time points in G₁ revealed that the requirement for MAPK/Erk activation lasts only to mid-G₁, while the actin CSK must remain intact up to late G₁ restriction point, R, in order for capillary endothelial cells to enter S phase. Additional analysis using Cyto D pulses defined a narrow time window of 3 h just prior to R in which CSK integrity was shown to be critical for the G₁/S transition. Cyto D treatment led to down-regulation of cyclin D1 protein and accumulation of the cdk inhibitor, p27^Kip1, independent of cell cycle phase, suggesting that these changes resulted directly from CSK disruption rather than from a general cell cycle block. Together, these data indicate the existence of a distinct time window in late G₁ in which signals elicited by the CSK act independently of early MAPK/Erk signals to drive the cell cycle machinery through the G₁/S boundary and, hence, promote cell growth.

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¹: To whom correspondence and reprint requests should be addressed at Departments of Pathology & Surgery, Children's Hospital and Harvard Medical School, Enders 1007, 300 Longwood Ave., Boston, MA 02115. Fax: 617-232-7914. E-mail: [email protected].

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Regular ArticleA Discrete Cell Cycle Checkpoint in Late G1 That Is Cytoskeleton-Dependent and MAP Kinase (Erk)-Independent

Abstract

Exp. Cell. Res.

Curr. Opin. Cell. Biol.

J. Biomechanics

Curr. Opin. Cell. Biol.

Cell

J. Biol. Chem.

J. Biol. Chem.

Curr. Biol.

Exp. Cell. Res.

J. Biol. Chem.

J. Biol. Chem.

Exp. Cell. Res.

J. Biol. Chem.

J. Biol. Chem.

Eur. J. Cell. Biol.

FEBS Lett.

Mol. Cell

J. Biol. Chem.

Biochem. Biophys. Res. Commun.

FEBS Lett.

Role of cell shape in growth control

Nature

Fibronectin controls capillary endothelial cell growth by modulating cell shape

Proc. Natl. Acad. Sci. USA

Cell-shape-dependent control of p27Kip and cell cycle progression in human capillary endothelial cells

Mol. Biol. Cell

The structural and mechanical complexity of cell-growth control Nat

Cell. Biol.

Actin cytoskeleton organization in response to integrin-mediated adhesion

Microsc. Res. Tech.

Integrin dynamics and matrix assembly: Tension-dependent translocation of alpha(5)beta(1) integrins promotes early fibronectin fibrillogenesis

J. Cell. Biol.

Growth regulation by cell shape alteration and organization of the cytoskeleton

Eur. J. Cell Biol.

Cytoskeletal integrity is required throughout the mitogen stimulation phase of the cell cycle and mediates the anchorage-dependent expression of cyclin D1

Mol. Biol. Cell.

Integrins and cell proliferation: Regulation of cyclin-dependent kinases via cytoplasmic signaling pathways

J. Cell. Sci.

Fibronectin, integrins, and growth control

J. Cell. Physiol.

G1 Phase progression: Cycling on cue

Cell

CDK inhibitors: Positive and negative regulators of G1-phase progression

Genes Dev.

G1 Events and regulation of cell proliferation

Science

Ras links growth factor signaling to the cell cycle machinery via regulation of cyclin D1 and the Cdk inhibitor p27KIP1

Mol. Cell Biol.

Regular Article
A Discrete Cell Cycle Checkpoint in Late G₁ That Is Cytoskeleton-Dependent and MAP Kinase (Erk)-Independent