Abnormalities of the large ribosomal subunit protein, Rpl35a, in Diamond-Blackfan anemia.
Diamond-Blackfan anemia (DBA) is an inherited bone marrow failure syndrome characterized by anemia, congenital abnormalities, and cancer predisposition. Small ribosomal subunit genes RPS19, RPS24, and RPS17 are mutated in approximately one-third of patients. We used a candidate gene strategy combining high-resolution genomic mapping and gene expression microarray in the analysis of 2 DBA patients with chromosome 3q deletions to identify RPL35A as a potential DBA gene. Sequence analysis of a cohort of DBA probands confirmed involvement RPL35A in DBA. shRNA inhibition shows that Rpl35a is essential for maturation of 28S and 5.8S rRNAs, 60S subunit biogenesis, normal proliferation, and cell survival. Analysis of pre-rRNA processing in primary DBA lymphoblastoid cell lines demonstrated similar alterations of large ribosomal subunit rRNA in both RPL35A-mutated and some RPL35A wild-type patients, suggesting additional large ribosomal subunit gene defects are likely present in some cases of DBA. These data demonstrate that alterations of large ribosomal subunit proteins cause DBA and support the hypothesis that DBA is primarily the result of altered ribosomal function. The results also establish that haploinsufficiency of large ribosomal subunit proteins contributes to bone marrow failure and potentially cancer predisposition.
Division of Pediatric Oncology, Department of Oncology, Kimmel Comprehensive Cancer Center.
Jason E
JE
Farrar
Michelle
M
Nater
Emi
E
Caywood
Michael A
MA
McDevitt
Jeanne
J
Kowalski
Clifford M
CM
Takemoto
C Conover
CC
Talbot
Paul
P
Meltzer
Diane
D
Esposito
Alan H
AH
Beggs
Hal E
HE
Schneider
Agnieszka
A
Grabowska
Sarah E
SE
Ball
Edyta
E
Niewiadomska
Colin A
CA
Sieff
Adrianna
A
Vlachos
Eva
E
Atsidaftos
Steven R
SR
Ellis
Jeffrey M
JM
Lipton
Hanna T
HT
Gazda
Robert J
RJ
Arceci
D000595:Amino Acid Sequence; D029503:Anemia, Diamond-Blackfan / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D016022:Case-Control Studies; D002460:Cell Line; D049109:Cell Proliferation; D002872:Chromosome Deletion; D002874:Chromosome Mapping; D002893:Chromosomes, Human, Pair 3 / Q000235:genetics; D015331:Cohort Studies; D004247:DNA / Q000235:genetics; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010375:Pedigree; D012323:RNA Processing, Post-Transcriptional; D034741:RNA, Small Interfering / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics* / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
D034741:RNA, Small Interfering; C489281:RPL35A protein, human; D012269:Ribosomal Proteins; D004247:DNA