title: Slow growth and unstable ribosomal RNA lacking pseudouridine in mouse embryonic fibroblast cells expressing catalytically inactive dyskerin.
abstract: Pseudouridine is the most abundant modified nucleotide in ribosomal RNA throughout eukaryotes and archaea but its role is not known. Here we produced mouse embryonic fibroblast cells expressing only catalytically inactive dyskerin, the pseudouridine synthase that converts uridine to pseudouridine in ribosomal RNA. The mutant dyskerin protein, D125A, was extremely unstable but cells were able to divide and grow very slowly. Abnormalities in ribosome RNA synthesis were apparent but mature cytoplasmic RNAs lacking pseudouridine were produced and were very unstable. We conclude that pseudouridine is required to stabilize the secondary structure of ribosomal RNA that is essential for its function.
authors.affiliation: Department of Pediatrics and Division of Hematology, The Children's Hospital of Philadelphia, 3615 Civic Center Boulevard, PA 19104-4318, USA.
authors.forename: Bai-Wei
authors.initials: BW
authors.lastname: Gu
authors.forename: Jingping
authors.initials: J
authors.lastname: Ge
authors.forename: Jian-Meng
authors.initials: JM
authors.lastname: Fan
authors.forename: Monica
authors.initials: M
authors.lastname: Bessler
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Mason
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology; D004795:Enzyme Stability; D005347:Fibroblasts / Q000201:enzymology* / Q000502:physiology; D051379:Mice; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D011560:Pseudouridine / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D020871:RNA Stability; D012335:RNA, Ribosomal / Q000378:metabolism*; D020537:RNA, Small Nucleolar / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; C510431:Dkc1 protein, mouse; D050505:Mutant Proteins; D009687:Nuclear Proteins; D012335:RNA, Ribosomal; D020537:RNA, Small Nucleolar; D011560:Pseudouridine