- Title
-
Ribosomal Protein Mutations Result in Constitutive p53 Protein Degradation through Impairment of the AKT Pathway
- Authors
- Antunes, A.T., Goos, Y.J., Pereboom, T.C., Hermkens, D., Wlodarski, M.W., Da Costa, L., MacInnes, A.W.
- Source
- Full text @ PLoS Genet.
Early cell death of RP mutant cells requires p53. A) Acridine orange staining allowing visualization of dying cells in wild type or rpS7 mutants at 1 dpf that are uninjected, injected with the p53 MO, or injected with the missense MO. Size bar = 0.25mm. B) Quantification of (A). **p<0.01. C) Acridine orange staining of a rpS7 mutant is observed predominantly in the brain region (arrowhead) or distributed over the surface of the tail (white boxes). D) Scoring results from o-dianisidine staining allowing visualization of hemoglobin-expressing cells in clutches of either 111 embryos (+mis MO) or 177 embryos (+p53 MO) from an rpS7 pairing. PHENOTYPE:
|
|
Caspase 3/7 activation is increasingly impaired in RP mutants. A) Caspase 2 and B) 3/7 activity is measured using a proluminescent caspase-3/7 DEVD-aminoluciferin substrate in 2 dpf embryos either untreated or exposed to 25 Gy ionizing radiation (IR). C) TUNEL analysis of 2 dpf wild type or rpS7 embryos reveals cells with DNA fragmentation when embryos are exposed to 25 Gy IR. D) The number of TUNEL-positive cells in (C) are quantified. E) TUNEL-positive cells in both wild type and rpS7 mutants are observed predominantly in the brain region (arrowhead) or in the tail tissue dorsal to the notochord (black box). *p<0.05, **p<0.01. PHENOTYPE:
|
|
RP mutations affect expression of βE1-globin but not scl A,B) In situ hybridizations of 2 dpf embryos measuring the mRNA expression levels of the transcription factor scl (A) and the globin gene βE1-globin (B). C) Representative shots of the dorsal aorta in either wild type of RP mutant embryos stained with probes against scl or βE1-globin. Note the decrease of the βE1-globin expression in the RP mutants compared to the wild type while the expression of scl remains unchanged. EXPRESSION / LABELING:
|
|
Morphology of zebrafish mutants with and without p53 MO. Light microscopy shots of representative wild type (left) or rpS7 mutant embryos either uninjected (center) or injected with the p53 MO (right) at 1, 2, or 3 dpf. Arrowheads indicate the inflation of the hindbrain vesicle and arrows indicate pericardial edemas, both phenotypes that are rescued by the p53 MO injection. PHENOTYPE:
|