Haploinsufficiency of Trp53 dramatically extends the lifespan of Sirt6-deficient mice
- The University of Hong Kong, Hong Kong
- Shenzhen University, China
- University of Rochester, United States
- Guangdong Medical University, China
Figures
Figure 1 with 1 supplement
Heterozygosity of Trp53 rescues premature senescence in Sirt6-deficient scenario, (Trp53 has been denoted as p53 in the figures).
(A) Quantification for qPCR analyses for gene expression of p53 targets (with respect to Gapdh controls) in Sirt6+/+Trp53+/+ (WT) and Sirt6-/-Trp53+/+ (Sirt6 KO) MEFs, respectively. Data represent …
Figure 1—figure supplement 1
Attenuation of p53 downstream targets and rescue of premature senescence-associated phenotypes in Sirt6-deficient scenario by haploinsufficiency of Trp53, (Trp53 has been denoted as p53 in the figures).
(A) Representative images of MEFs derived from Sirt6+/+Trp53+/+, Sirt6-/-Trp53+/+ and Sirt6-/-Trp53+/- embryos at P6. Scale bar, 100 µm. (B) qPCR analyses of p53 mRNA levels in Sirt6+/+Trp53+/+, …
Figure 2 with 1 supplement
Haploinsufficiency of Trp53 significantly extends the lifespan of Sirt6-deficient mice, (Trp53 has been denoted as p53 in the figures).
(A) Representative images of Sirt6+/+Trp53+/+(WT), Sirt6-/-Trp53+/+ (Sirt6 KO), and -/-Trp53+/- (compound mutant) littermate mice at the age of 24 days. (B) Mean body weights of WT, Sirt6 KO and …
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Figure 2—source data 1
Serum Igf-1 levels in 24-day old mice.
- https://doi.org/10.7554/eLife.32127.007
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Figure 2—source data 2
Serum Igf-1 levels in 10-16 months old mice.
- https://doi.org/10.7554/eLife.32127.008
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Figure 2—source data 3
Serum glucose levels in 24-day old mice.
- https://doi.org/10.7554/eLife.32127.009
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Figure 2—source data 4
Serum glucose levels in 10-16 months old mice.
- https://doi.org/10.7554/eLife.32127.010
Figure 2—figure supplement 1
Analysis of serum IGF-1 and glucose levels in Sirt6+/+Trp53+/+, Sirt6-/-Trp53+/+ and Sirt6-/-Trp53+/- mice, (Trp53 has been denoted as p53 in the figures).
(A) Serum IGF-1 levels in Sirt6+/+Trp53+/+, Sirt6-/-Trp53+/+ and Sirt6-/-Trp53+/- mice at around 24 days. P value calculated using one-way ANOVA. (B) Serum IGF-1 levels in Sirt6+/+Trp53+/+ and Sirt6-…
Figure 3 with 1 supplement
Amelioration of premature-aging associated phenotypes in Sirt6 KO mice upon haploinsufficiency of p53, (Trp53 has been denoted as p53 in the figures).
(A) Representative images of spleens, thymus, livers, kidneys, lungs and hearts from 24 days old Sirt6+/+Trp53+/+, Sirt6-/-Trp53+/+, and Sirt6-/-Trp53+/- mice. (B) Whole body X-ray images of female …
Figure 3—figure supplement 1
Incidence of cancer and other age-associated disorders in Sirt6-/-Trp53+/- mice, (Trp53 has been denoted as p53 in the figures).
(A) Whole body X-ray images of female and male Sirt6+/+Trp53+/+, Sirt6-/-Trp53+/+ and Sirt6-/-Trp53+/- mice (note the lordokyphosis/curved spine in Sirt6-/-Trp53+/+ mice) at the age of 24 days. (B) …
Figure 4
Haploinsufficiency of Trp53 prevents bone marrow stem cell decline, suppresses apoptosis, and restores immune cell populations in Sirt6-deficient mice, (Trp53 has been denoted as p53 in the figures).
(A) Percentage of mesenchymal stem cell (MSC) population in the female Sirt6+/+Trp53+/+, Sirt6-/-Trp53+/+ and Sirt6-/-Trp53+/- mice at the age of ~24 days. Data represent mean ± SEM; n=5. (B) …
Figure 5 with 1 supplement
SIRT6 directly deacetylates p53 at lysine 381, but not K382.
(A) Western blotting analysis of endogenous interaction between SIRT6 and p53 in mock CRISPR control cells and SIRT6 KO cells using specific antibodies for SIRT6 and p53, keeping respective IgG …
Figure 5—figure supplement 1
Analyses of physical and functional interaction between SIRT6 and p53, (Trp53 has been denoted as p53 in the figures).
(A) Western blotting analysis of endogenous p53 pulled down by FLAG-tagged SIRT6 in HEK293 cells. (B) Western blotting analysis of endogenous SIRT6 pulled down by FLAG-tagged p53 in HEK293 cells. (C)…
Figure 6 with 2 supplements
SIRT6 negatively regulates the stability of p53.
(A) Western blotting analysis of p53 protein expression in SIRT6 KO and control HEK293 cells. (B) Quantification of data presented in (A). Data represent mean ± SEM, n = 3. **p<0.01 calculated using …
Figure 6—figure supplement 1
Analyses of the stability of p53 in the presence and absence of SIRT6.
(A) Western blotting analysis of p53 protein expression in Sirt6+/+ and Sirt6-/- MEFs. (B) Immunofluorescence staining to confirm enhanced p53 protein expression in WT and Sirt6-/- MEFs. Scale bar, …
Figure 6—figure supplement 2
Acetyl mimic mutant p53 K381Q confers stability to p53 and imparts senescence-like properties to cells.
(A) FLAG-tagged p53 wild-type (WT), lysine 381 to arginine (K381R, non-acetylatable) mutant, and lysine 381 to glutamine (K381Q, acetyl-mimic) mutants were ectopically expressed in HEK293 cells. …
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.32127.019
