6% depending on tumor type), with pathogenicity varying from benign to deleterious by in

silico predictions. At least one colon Selleckchem Afatinib cancer case with a somatic missense change (R79C) is included. 6 Tumors from mutation carriers showed no loss of the wild-type allele (Supplementary Figure 2B), arguing against Knudson’s 2-hit mechanism for tumor-suppressor genes. 7 The absence of loss of heterozygosity complies with observations from zebrafish showing that ribosomal protein genes act as haploinsufficient suppressors of tumorigenesis. 8 RPS20 is required during the late steps of 18S ribosomal RNA (rRNA) formation.9 Indeed, Northern blot analysis showed that small interfering RNA depletion of RPS20 in HeLa cells led to a significant increase of 21S pre-rRNAs (which are distributed in 2 close bands in this cell type), as well as an accumulation of 18S-E pre-rRNAs (Figure 2A). This was accompanied by a strong decrease of the 18S/28S ratio ( Figure 2B). Patients

carrying the RPS20 c.147dupA mutation (A1–A4) showed a marked increase of 21S pre-rRNAs compared with healthy unrelated controls (C1–C3), while the Pictilisib in vivo 18S-E pre-rRNA level was in the same range in control, noncarrier, and patient samples ( Figure 2C). The 18S/28S ratios were unchanged in patient cells compared with controls and a noncarrier. Altogether, these results show a late pre-rRNA processing defect in mutation carrier cells consistent with RPS20 haploinsufficiency. Polysome analysis showed a slight increase in the 60S peak relative to the

40S peak in mutation carriers compared with a noncarrier and a healthy unrelated control ( Supplementary Figure 3). Collectively, Buspirone HCl RNA results suggest that the RPS20 mutation disturbs ribosome biogenesis by affecting the equilibrium between the different pre-rRNA species and the formation of mature 18S rRNA. All RPSs are essential in human cells, except RPS25.9 The ribosomal protein gene family comprises 80 genes,8 at least 11 of which are known to be mutated in Diamond–Blackfan anemia, a dominantly inherited form of pure red cell aplasia, growth retardation, and congenital anomalies.10 and 11 No such features were present in colon cancer patients from F56. Why is the RPS20 mutation associated with colorectal cancer susceptibility, while mutations in 11 other ribosomal protein genes cause predisposition to Diamond–Blackfan anemia? Haploinsufficiency for RPS19 or RPS20 in mice was shown to stabilize p53, which in turn had different effects in different cell types. 12 Mouse findings make it tempting to speculate that cell type–specific effects of RPS20 haploinsufficiency might play a role in RPS20-associated colon tumorigenesis in human beings, with disturbed ribosome biogenesis, altered p53 dosage, or various downstream events as possible mediators. Among ribosomal proteins, “detector” and “effector” types have been distinguished based on contribution to p53 stress response.