Ribosomal protein S27-like is a physiological regulator of p53 that suppresses genomic instability and tumorigenesis
- University of Michigan, United States
- Children's National Medical Center, United States
- University of Michigan Medical School, United States
- Zhejiang University School of Medicine, China
Figures
Figure 1 with 1 supplement
Rps27l disruption causes postnatal death as a result of increased apoptosis.
(A) Disruption of Rps27l causes postnatal death. (B–D) Reduced body size, body weight, and organ weight in Rps27l−/− mice. An Rps27l−/− pup (bottom) and two Rps27l+/− littermates (top) at P8 were …
Figure 1—figure supplement 1
Generation of Rps27l gene trap mice and phenotypes of Rps27l−/− mice.
(A) Genomic structure of mouse Rps27l gene. Four exons (E1-E4) and three introns (between exons) are shown, along with the insertion site in intron 1 of the gene trap vector. (B) Southern blotting …
Figure 2 with 1 supplement
Reduced hematopoietic stem and progenitor cells in Rps27l−/− fetal livers.
(A and B) Reduced size and total cell numbers in Rps27l−/− fetal livers. Representative fetal livers of three genotypes were photographed (A), and the numbers of fetal liver cells from E14.5 embryos …
Figure 2—figure supplement 1
Depletion of hematopoietic stem and progenitor cells (HSPCs) upon Rps27l disruption.
(A) Representative FACS profiles of HSPCs in fetal livers. Fetal liver cells were isolated from E14.5 embryos with indicated genotypes and subjected to FACS analysis using indicated surface marker …
Figure 3 with 1 supplement
Rps27l disruption causes a moderate increase of p53 and p53 targets.
(A) Representative p53 staining in fetal livers and bone marrows. Fetal livers at E14.5 and bone marrows at P6 were immuno-stained with p53 Ab. Arrows point to p53 positive staining. Scale bar …
Figure 3—figure supplement 1
Increased p53 levels in responsive to various stresses in Rps27l−/− MEFs.
(A) Higher p53 levels induced by ribosomal stress and DNA damaging agent, not by endoplasmic reticulum (ER) stress in Rps27l−/− MEFs. MEFs were treated with 5 nM Actinomycin D (ActD) for 24 hr, 5 …
Figure 4 with 1 supplement
Rps27l disruption alters the levels of the p53-Mdm2-Mdm4 axis.
(A) Extension of protein half-lives of p53 and Mdm2 upon Rps27l disruption. Rps27l+/+ or Rps27l−/− MEFs were harvested at various time points post CHX treatment for IB (top). Densitometry …
Figure 4—figure supplement 1
Rps27l regulates p53 ubiquitylation and Mdm2 protein half-life.
(A) No change in p53 transcription, but an increase in transcription of p53 targets. Four independent pairs of MEFs were harvested for quantitative real-time PCR using indicated primers. Shown are …
Figure 5 with 1 supplement
Rps27l is a ribosomal protein whose depletion triggers ribosomal stress.
(A) Rps27l is a ribosomal protein. Rps27l+/+ or Rps27l−/− MEFs were treated with CHX for 30 min before harvesting for ribosomal profiling. The cytoplasmic extracts were loaded on sucrose gradients …
Figure 5—figure supplement 1
Rps27l disruption has minimal effects on rDNA transcription, rRNA processing and/or protein synthesis.
(A) Rps27l loss does not impair the transcription of rDNA. Four independent pairs of MEFs were harvested for quantitative real-time PCR using primers for 45S rRNA. Shown are mean ± SEM (n = 4). (B) …
Figure 6 with 1 supplement
Rps27l depletion leads to Mdm2-mediated Mdm4 degradation.
(A) Increased Mdm4 ubiquitylation upon Rps27l disruption. Rps27l+/+ or Rps27l−/− MEFs were treated with MG132 before being harvested for IP with Mdm4 Ab or normal IgG control, followed by IB with …
Figure 6—figure supplement 1
Rps27l regulates Mdm2-mediated Mdm4 degradation.
(A) No change in Mdm4 mRNA transcription. Four independent pairs of MEFs were harvested for qRT-PCR using indicated primers. Shown are mean ± SEM (n = 2). (B) Reduced Mdm4 protein level in Rps27l−/− …
Figure 7 with 1 supplement
Simultaneous deletion of Trp53 rescues growth retardation and HSPCs depletion.
(A) Deletion of Trp53 rescues postnatal death by Rps27l disruption. Lower than expected number of mice with Trp53−/− genotype (regardless of Rps27l genotype) is due to high frequency of …
Figure 7—figure supplement 1
Simultaneous deletion of Trp53 rescues defective phenotypes caused by Rps27l disruption.
(A and B) Significant HSPCs decrease in bone marrow of Rps27l−/− mice. Bone marrow cells from Rps27l+/+ (n = 4), Rps27l+/− (n = 4), and Rps27l−/− (n = 5) mice at P18 under a mixed Sv129/B6 …
Figure 8 with 1 supplement
Rps27l disruption induces genomic instability and spontaneous lymphoma.
(A and B) Kaplan–Meier survival curves of Trp53−/− (A) and Trp53+/− (B) mice with three Rps27l genotypes and indicated numbers of mice. p = 0.713 (A); p = 0.0036 (B). (C) Representative pictures of …
Figure 8—figure supplement 1
Rps27l disruption induces genomic instability and spontaneous lymphoma.
(A) Representative staining with H&E and CD3, a T cell marker of indicated tissues from a Rps27l−/−;Trp53+/− mouse. Scale bar = 50 µm. (B) Representative staining with H&E and B220, a B cell marker …
Figure 9
Rps27l is a p53 regulator and also a p53 ‘goalkeeper’—a working model.
In normal cell with wild-type p53 (Trp53+/+), Rps27l, upon induction by p53 in response to various stresses, stabilizes the Mdm2-Mdm4 heterodimer to form an optimal E3 ligase complex for effective …
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