Ribosomal protein S19 (RPS19) is frequently mutated in Diamond-Blackfan anemia 
(DBA), a rare congenital hypoplastic anemia. Recent studies have shown that 
RPS19 expression decreases during terminal erythroid differentiation. Currently 
no information is available on the subcellular localization of normal RPS19 and 
the potential effects of various RPS19 mutations on cellular localization. In 
the present study, using wild-type and mutant RPS19 cDNA, we explored the 
subcellular distribution of normal and mutant proteins in a fibroblast cell line 
(Cos-7 cells). RPS19 was detected primarily in the nucleus, and more 
specifically in the nucleoli, where RPS19 colocalized with the nucleolar protein 
nucleolin. Using various N-terminal and C-terminal deletion constructs, we 
identified 2 nucleolar localization signals (NoSs) in RPS19: the first 
comprising amino acids Met1 to Arg16 in the NH2-terminus and the second 
comprising Gly120 to Asn142 in the COOH-terminus. Importantly, 2 mutations 
identified in DBA patients, Val15Phe and Gly127Gln, each of which localized to 1 
of the 2 NoS, failed to localize RPS19 to the nucleolus. In addition to their 
mislocalization, there was a dramatic decrease in the expression of the 2 mutant 
proteins compared to the wild type. This decrease in protein expression was 
specific for the mutant RPS19, since expression of other proteins was normal. 
The present findings enable us to document the nucleolar localization signals in 
RPS19 and help define the phenotypic consequences of some mutations in RPS19 in 
DBA.