Perturbation of rRNA synthesis in the bap28 mutation leads to apoptosis mediated by p53 in the zebrafish central nervous system.
Zebrafish is a powerful vertebrate model system for using forward genetics to elucidate mechanisms of early development. We have used chemical mutagenesis to screen for mutants that show defects in the CNS. Here we describe the isolation of the bap28 mutation that leads to abnormalities in the brain starting at midsomitogenesis stages. Mutant embryos display excess apoptosis primarily in the central nervous system (CNS) and die by days 6-7 after fertilization. The mutation was positionally cloned and shown to affect a gene that encodes a large protein with high similarity to the uncharacterized human protein BAP28 and lower similarity to yeast Utp10. Utp10 is a component of a nucleolar U3 small nucleolar RNA-containing RNP complex that is required for transcription of ribosomal DNA and for processing of 18 S rRNA. We show that zebrafish Bap28 likewise is required for rRNA transcription and processing, with a major effect on 18 S rRNA maturation. We suggest that bap28 is required for cell survival in the CNS through its role in rRNA synthesis and processing. Inhibition of p53 protein expression in bap28 mutants led to embryos with morphologically normal appearance, suggesting that p53 is involved in triggering apoptosis in the bap28 mutant CNS. The bap28 mutation provides a genetic approach to study the role of ribosome biogenesis in the development of a vertebrate embryo.
Azuma
Mizuki
M
Laboratory of Molecular Genetics, NICHD, National Institutes of Health, Bethesda, Maryland 20892.
Toyama
Reiko
R
Laboratory of Molecular Genetics, NICHD, National Institutes of Health, Bethesda, Maryland 20892.
Laver
Elizabeth
E
Laboratory of Molecular Genetics, NICHD, National Institutes of Health, Bethesda, Maryland 20892.
Dawid
Igor B
IB
Laboratory of Molecular Genetics, NICHD, National Institutes of Health, Bethesda, Maryland 20892. Electronic address: idawid@nih.gov.
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D001921:Brain / Q000254:growth & development / Q000378:metabolism; D002490:Central Nervous System / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D009154:Mutation; D012335:RNA, Ribosomal / Q000096:biosynthesis*; D012337:RNA, Ribosomal, 18S / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
D012335:RNA, Ribosomal; D012337:RNA, Ribosomal, 18S; D016159:Tumor Suppressor Protein p53; D029961:Zebrafish Proteins; C511611:heatr1 protein, zebrafish