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Chronic Replication Problems Impact Cell Morphology and Adhesion of DNA Ligase I Defective Cells.

Cremaschi P, Oliverio M, Leva V, Bione S, Carriero R, Mazzucco G, Palamidessi A, Scita G, Biamonti G, Montecucco A - PLoS ONE (2015)

Bottom Line: This replication stress elicits a constitutive phosphorylation of the ataxia telangiectasia mutated (ATM) checkpoint kinase that fails to arrest cell cycle progression or to activate apoptosis or cell senescence.Here we show that parental 46BR.1G1 and 7A3 cells differ in important features such as cell morphology, adhesion and migration.Interestingly, ATM inhibition makes 46BR.1G1 more similar to 7A3 cells for what concerns morphology, adhesion and expression of cell-cell adhesion receptors.

View Article: PubMed Central - PubMed

Affiliation: Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche (CNR), Pavia, Italy; Dipartimento di Biologia e Biotecnologie "L. Spallanzani", Università degli Studi di Pavia, Pavia, Italy.

ABSTRACT
Moderate DNA damage resulting from metabolic activities or sub-lethal doses of exogenous insults may eventually lead to cancer onset. Human 46BR.1G1 cells bear a mutation in replicative DNA ligase I (LigI) which results in low levels of replication-dependent DNA damage. This replication stress elicits a constitutive phosphorylation of the ataxia telangiectasia mutated (ATM) checkpoint kinase that fails to arrest cell cycle progression or to activate apoptosis or cell senescence. Stable transfection of wild type LigI, as in 7A3 cells, prevents DNA damage and ATM activation. Here we show that parental 46BR.1G1 and 7A3 cells differ in important features such as cell morphology, adhesion and migration. Comparison of gene expression profiles in the two cell lines detects Bio-Functional categories consistent with the morphological and migration properties of LigI deficient cells. Interestingly, ATM inhibition makes 46BR.1G1 more similar to 7A3 cells for what concerns morphology, adhesion and expression of cell-cell adhesion receptors. These observations extend the influence of the DNA damage response checkpoint pathways and unveil a role for ATM kinase activity in modulating cell biology parameters relevant to cancer progression.

No MeSH data available.


Related in: MedlinePlus

LigI-deficient 46BR.1G1 cells adhere more efficiently to the plate than complemented 7A3 cells.Cells were plated on 96-well plate and allowed to adhere for 30 minutes before fixing. Cells were stained with Crystal Violet, solubilized with acetic acid and quantified by measuring the OD at 620 nm. Data are shown as mean ± SEM of four independent experiments.
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pone.0130561.g002: LigI-deficient 46BR.1G1 cells adhere more efficiently to the plate than complemented 7A3 cells.Cells were plated on 96-well plate and allowed to adhere for 30 minutes before fixing. Cells were stained with Crystal Violet, solubilized with acetic acid and quantified by measuring the OD at 620 nm. Data are shown as mean ± SEM of four independent experiments.

Mentions: Changes in cell morphology may be linked to an altered cell adhesion. To verify this aspect, we challenged the two cell lines in a standard cell adhesion assay. As shown in Fig 2, 46BR.1G1 cells adhered more efficiently to the plate than LigI-proficient 7A3 cells. Notably, incubation with caffeine and KU-55933 significantly reduced adhesion of 46BR.1G1 but not of 7A3 cells. Altogether these results suggest that the activation of the ATM/Chk2 signaling pathway has an important role in the effect of replication stress induced by LigI-deficiency on cytoskeleton organization and cell adhesiveness.


Chronic Replication Problems Impact Cell Morphology and Adhesion of DNA Ligase I Defective Cells.

Cremaschi P, Oliverio M, Leva V, Bione S, Carriero R, Mazzucco G, Palamidessi A, Scita G, Biamonti G, Montecucco A - PLoS ONE (2015)

LigI-deficient 46BR.1G1 cells adhere more efficiently to the plate than complemented 7A3 cells.Cells were plated on 96-well plate and allowed to adhere for 30 minutes before fixing. Cells were stained with Crystal Violet, solubilized with acetic acid and quantified by measuring the OD at 620 nm. Data are shown as mean ± SEM of four independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495043&req=5

pone.0130561.g002: LigI-deficient 46BR.1G1 cells adhere more efficiently to the plate than complemented 7A3 cells.Cells were plated on 96-well plate and allowed to adhere for 30 minutes before fixing. Cells were stained with Crystal Violet, solubilized with acetic acid and quantified by measuring the OD at 620 nm. Data are shown as mean ± SEM of four independent experiments.
Mentions: Changes in cell morphology may be linked to an altered cell adhesion. To verify this aspect, we challenged the two cell lines in a standard cell adhesion assay. As shown in Fig 2, 46BR.1G1 cells adhered more efficiently to the plate than LigI-proficient 7A3 cells. Notably, incubation with caffeine and KU-55933 significantly reduced adhesion of 46BR.1G1 but not of 7A3 cells. Altogether these results suggest that the activation of the ATM/Chk2 signaling pathway has an important role in the effect of replication stress induced by LigI-deficiency on cytoskeleton organization and cell adhesiveness.

Bottom Line: This replication stress elicits a constitutive phosphorylation of the ataxia telangiectasia mutated (ATM) checkpoint kinase that fails to arrest cell cycle progression or to activate apoptosis or cell senescence.Here we show that parental 46BR.1G1 and 7A3 cells differ in important features such as cell morphology, adhesion and migration.Interestingly, ATM inhibition makes 46BR.1G1 more similar to 7A3 cells for what concerns morphology, adhesion and expression of cell-cell adhesion receptors.

View Article: PubMed Central - PubMed

Affiliation: Istituto di Genetica Molecolare, Consiglio Nazionale delle Ricerche (CNR), Pavia, Italy; Dipartimento di Biologia e Biotecnologie "L. Spallanzani", Università degli Studi di Pavia, Pavia, Italy.

ABSTRACT
Moderate DNA damage resulting from metabolic activities or sub-lethal doses of exogenous insults may eventually lead to cancer onset. Human 46BR.1G1 cells bear a mutation in replicative DNA ligase I (LigI) which results in low levels of replication-dependent DNA damage. This replication stress elicits a constitutive phosphorylation of the ataxia telangiectasia mutated (ATM) checkpoint kinase that fails to arrest cell cycle progression or to activate apoptosis or cell senescence. Stable transfection of wild type LigI, as in 7A3 cells, prevents DNA damage and ATM activation. Here we show that parental 46BR.1G1 and 7A3 cells differ in important features such as cell morphology, adhesion and migration. Comparison of gene expression profiles in the two cell lines detects Bio-Functional categories consistent with the morphological and migration properties of LigI deficient cells. Interestingly, ATM inhibition makes 46BR.1G1 more similar to 7A3 cells for what concerns morphology, adhesion and expression of cell-cell adhesion receptors. These observations extend the influence of the DNA damage response checkpoint pathways and unveil a role for ATM kinase activity in modulating cell biology parameters relevant to cancer progression.

No MeSH data available.


Related in: MedlinePlus