title: Exploiting pre-rRNA processing in Diamond Blackfan anemia gene discovery and diagnosis.
abstract: Diamond Blackfan anemia (DBA), a syndrome primarily characterized by anemia and physical abnormalities, is one among a group of related inherited bone marrow failure syndromes (IBMFS) which share overlapping clinical features. Heterozygous mutations or single-copy deletions have been identified in 12 ribosomal protein genes in approximately 60% of DBA cases, with the genetic etiology unexplained in most remaining patients. Unlike many IBMFS, for which functional screening assays complement clinical and genetic findings, suspected DBA in the absence of typical alterations of the known genes must frequently be diagnosed after exclusion of other IBMFS. We report here a novel deletion in a child that presented such a diagnostic challenge and prompted development of a novel functional assay that can assist in the diagnosis of a significant fraction of patients with DBA. The ribosomal proteins affected in DBA are required for pre-rRNA processing, a process which can be interrogated to monitor steps in the maturation of 40S and 60S ribosomal subunits. In contrast to prior methods used to assess pre-rRNA processing, the assay reported here, based on capillary electrophoresis measurement of the maturation of rRNA in pre-60S ribosomal subunits, would be readily amenable to use in diagnostic laboratories. In addition to utility as a diagnostic tool, we applied this technique to gene discovery in DBA, resulting in the identification of RPL31 as a novel DBA gene.
authors.affiliation: Pediatric Hematology/Oncology Section, Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, Arkansas.
authors.forename: Jason E
authors.initials: JE
authors.lastname: Farrar
authors.forename: Paola
authors.initials: P
authors.lastname: Quarello
authors.forename: Ross
authors.initials: R
authors.lastname: Fisher
authors.forename: Kelly A
authors.initials: KA
authors.lastname: O'Brien
authors.forename: Anna
authors.initials: A
authors.lastname: Aspesi
authors.forename: Sara
authors.initials: S
authors.lastname: Parrella
authors.forename: Adrianna L
authors.initials: AL
authors.lastname: Henson
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Seidel
authors.forename: Eva
authors.initials: E
authors.lastname: Atsidaftos
authors.forename: Supraja
authors.initials: S
authors.lastname: Prakash
authors.forename: Shahla
authors.initials: S
authors.lastname: Bari
authors.forename: Emanuela
authors.initials: E
authors.lastname: Garelli
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Arceci
authors.forename: Irma
authors.initials: I
authors.lastname: Dianzani
authors.forename: Ugo
authors.initials: U
authors.lastname: Ramenghi
authors.forename: Adrianna
authors.initials: A
authors.lastname: Vlachos
authors.forename: Jeffrey M
authors.initials: JM
authors.lastname: Lipton
authors.forename: David M
authors.initials: DM
authors.lastname: Bodine
authors.forename: Steven R
authors.initials: SR
authors.lastname: Ellis
mesh_terms: D029503:Anemia, Diamond-Blackfan / Q000235:genetics / Q000378:metabolism; D005260:Female; D006801:Humans; D007223:Infant; D020014:K562 Cells; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012335:RNA, Ribosomal / Q000378:metabolism; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism; D054683:Ribosome Subunits, Large, Eukaryotic / Q000235:genetics / Q000378:metabolism; D054682:Ribosome Subunits, Small, Eukaryotic / Q000235:genetics / Q000378:metabolism
chemical_list: D012322:RNA Precursors; D012335:RNA, Ribosomal; D012269:Ribosomal Proteins; C041672:ribosomal protein L34