Limits...
p53 protects against genome instability following centriole duplication failure.

Lambrus BG, Uetake Y, Clutario KM, Daggubati V, Snyder M, Sluder G, Holland AJ - J. Cell Biol. (2015)

Bottom Line: Depleting p53 allowed cells that fail centriole duplication to proliferate indefinitely.Washout of auxin and restoration of endogenous Plk4 levels in cells that lack centrioles led to the penetrant formation of de novo centrioles that gained the ability to organize microtubules and duplicate.In summary, we uncover a p53-dependent surveillance mechanism that protects against genome instability by preventing cell growth after centriole duplication failure.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.

Show MeSH

Related in: MedlinePlus

The IAA-induced cell cycle arrest is not caused by a prolonged mitosis. (A) Representative images of anaphase cells at indicated times after IAA addition. Cells were costained with Centrin and CEP192. Bars: (main) 5 µm; (inset) 0.5 µm. (B) Quantification of the percentage of cell divisions occurring with the indicated number of centrioles at each spindle pole. Measurements were obtained from fixed samples at the indicated times after IAA addition. Bars represent the mean of >60 cells from two independent experiments. (C) Schematic showing the expected dynamics of centriole dilution after IAA treatment. Cells that undergo mitosis within 6 h of IAA addition almost always contain four centrioles, which dilute out as shown in subsequent divisions. (D) Graph showing the prometaphase duration and proliferative capacity of IAA-treated Plk4AID/AID cells. Each bar represents a daughter cell, its height represents the prometaphase duration of the mother cell, and its color represents the fate of the daughter. Only cells that underwent their first mitosis within 6 h of IAA treatment were analyzed. The dashed red line indicates the maximum time that IAA-treated cells spend in prometaphase before undergoing a cell cycle arrest (Fig. S3 D). Data are taken from two independent experiments (n = 199 prometaphases). (E) Immunoblot showing the levels of phosphorylated LATS, YAP, and Histone H2AX in IAA-treated Plk4AID/AID and parental RPE1 cells. Doxorubicin treatment was used as a control to induce DNA damage. (F) Graph showing the fold increase in cell number in IAA-treated Plk4AID/AID cells grown in normal (21%) or low (3%) oxygen conditions. Points show the mean of two independent experiments performed in triplicate. All error bars in the figure represent the SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4494000&req=5

fig4: The IAA-induced cell cycle arrest is not caused by a prolonged mitosis. (A) Representative images of anaphase cells at indicated times after IAA addition. Cells were costained with Centrin and CEP192. Bars: (main) 5 µm; (inset) 0.5 µm. (B) Quantification of the percentage of cell divisions occurring with the indicated number of centrioles at each spindle pole. Measurements were obtained from fixed samples at the indicated times after IAA addition. Bars represent the mean of >60 cells from two independent experiments. (C) Schematic showing the expected dynamics of centriole dilution after IAA treatment. Cells that undergo mitosis within 6 h of IAA addition almost always contain four centrioles, which dilute out as shown in subsequent divisions. (D) Graph showing the prometaphase duration and proliferative capacity of IAA-treated Plk4AID/AID cells. Each bar represents a daughter cell, its height represents the prometaphase duration of the mother cell, and its color represents the fate of the daughter. Only cells that underwent their first mitosis within 6 h of IAA treatment were analyzed. The dashed red line indicates the maximum time that IAA-treated cells spend in prometaphase before undergoing a cell cycle arrest (Fig. S3 D). Data are taken from two independent experiments (n = 199 prometaphases). (E) Immunoblot showing the levels of phosphorylated LATS, YAP, and Histone H2AX in IAA-treated Plk4AID/AID and parental RPE1 cells. Doxorubicin treatment was used as a control to induce DNA damage. (F) Graph showing the fold increase in cell number in IAA-treated Plk4AID/AID cells grown in normal (21%) or low (3%) oxygen conditions. Points show the mean of two independent experiments performed in triplicate. All error bars in the figure represent the SEM.

Mentions: Using fixed imaging, we examined the number of centrioles at various time points after IAA treatment. As expected, untreated control cells divided with two centrioles at each spindle pole (Fig. 4, A and B). In contrast, at 1 d after IAA addition, >90% of Plk4AID/AID cells divided with a single centriole at each spindle pole, whereas at days 2 and 3 after IAA addition >65% of cells divided with an asymmetric spindle comprised of an acentriolar pole and a pole containing a single centriole (Fig. 4, A and B). Cell divisions were rarely observed to take place in the absence of centrioles.


p53 protects against genome instability following centriole duplication failure.

Lambrus BG, Uetake Y, Clutario KM, Daggubati V, Snyder M, Sluder G, Holland AJ - J. Cell Biol. (2015)

The IAA-induced cell cycle arrest is not caused by a prolonged mitosis. (A) Representative images of anaphase cells at indicated times after IAA addition. Cells were costained with Centrin and CEP192. Bars: (main) 5 µm; (inset) 0.5 µm. (B) Quantification of the percentage of cell divisions occurring with the indicated number of centrioles at each spindle pole. Measurements were obtained from fixed samples at the indicated times after IAA addition. Bars represent the mean of >60 cells from two independent experiments. (C) Schematic showing the expected dynamics of centriole dilution after IAA treatment. Cells that undergo mitosis within 6 h of IAA addition almost always contain four centrioles, which dilute out as shown in subsequent divisions. (D) Graph showing the prometaphase duration and proliferative capacity of IAA-treated Plk4AID/AID cells. Each bar represents a daughter cell, its height represents the prometaphase duration of the mother cell, and its color represents the fate of the daughter. Only cells that underwent their first mitosis within 6 h of IAA treatment were analyzed. The dashed red line indicates the maximum time that IAA-treated cells spend in prometaphase before undergoing a cell cycle arrest (Fig. S3 D). Data are taken from two independent experiments (n = 199 prometaphases). (E) Immunoblot showing the levels of phosphorylated LATS, YAP, and Histone H2AX in IAA-treated Plk4AID/AID and parental RPE1 cells. Doxorubicin treatment was used as a control to induce DNA damage. (F) Graph showing the fold increase in cell number in IAA-treated Plk4AID/AID cells grown in normal (21%) or low (3%) oxygen conditions. Points show the mean of two independent experiments performed in triplicate. All error bars in the figure represent the SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4494000&req=5

fig4: The IAA-induced cell cycle arrest is not caused by a prolonged mitosis. (A) Representative images of anaphase cells at indicated times after IAA addition. Cells were costained with Centrin and CEP192. Bars: (main) 5 µm; (inset) 0.5 µm. (B) Quantification of the percentage of cell divisions occurring with the indicated number of centrioles at each spindle pole. Measurements were obtained from fixed samples at the indicated times after IAA addition. Bars represent the mean of >60 cells from two independent experiments. (C) Schematic showing the expected dynamics of centriole dilution after IAA treatment. Cells that undergo mitosis within 6 h of IAA addition almost always contain four centrioles, which dilute out as shown in subsequent divisions. (D) Graph showing the prometaphase duration and proliferative capacity of IAA-treated Plk4AID/AID cells. Each bar represents a daughter cell, its height represents the prometaphase duration of the mother cell, and its color represents the fate of the daughter. Only cells that underwent their first mitosis within 6 h of IAA treatment were analyzed. The dashed red line indicates the maximum time that IAA-treated cells spend in prometaphase before undergoing a cell cycle arrest (Fig. S3 D). Data are taken from two independent experiments (n = 199 prometaphases). (E) Immunoblot showing the levels of phosphorylated LATS, YAP, and Histone H2AX in IAA-treated Plk4AID/AID and parental RPE1 cells. Doxorubicin treatment was used as a control to induce DNA damage. (F) Graph showing the fold increase in cell number in IAA-treated Plk4AID/AID cells grown in normal (21%) or low (3%) oxygen conditions. Points show the mean of two independent experiments performed in triplicate. All error bars in the figure represent the SEM.
Mentions: Using fixed imaging, we examined the number of centrioles at various time points after IAA treatment. As expected, untreated control cells divided with two centrioles at each spindle pole (Fig. 4, A and B). In contrast, at 1 d after IAA addition, >90% of Plk4AID/AID cells divided with a single centriole at each spindle pole, whereas at days 2 and 3 after IAA addition >65% of cells divided with an asymmetric spindle comprised of an acentriolar pole and a pole containing a single centriole (Fig. 4, A and B). Cell divisions were rarely observed to take place in the absence of centrioles.

Bottom Line: Depleting p53 allowed cells that fail centriole duplication to proliferate indefinitely.Washout of auxin and restoration of endogenous Plk4 levels in cells that lack centrioles led to the penetrant formation of de novo centrioles that gained the ability to organize microtubules and duplicate.In summary, we uncover a p53-dependent surveillance mechanism that protects against genome instability by preventing cell growth after centriole duplication failure.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205.

Show MeSH
Related in: MedlinePlus