Cited by 27Nuclear Structure and FunctionRNA and protein synthesis mechanismsRNA Research and Splicing

Deletion of drosophila nopp140 induces subcellular ribosomopathies

He, Fang, James, Allison, Raje, Himanshu, Ghaffari, Helya, DiMario, Patrick·2015·Chromosoma

Abstract

The nucleolar and Cajal body phosphoprotein of 140 kDa (Nopp140) is considered a ribosome assembly factor, but its precise functions remain unknown. To approach this problem, we deleted the Nopp140 gene in Drosophila using FLP-FRT recombination. Genomic PCR, reverse transcriptase-PCR (RT-PCR), and immunofluorescence microscopy confirmed the loss of Nopp140, its messenger RNA (mRNA), and protein products from all tissues examined. Nopp140-/- larvae arrested in the second instar stage and most died within 8 days. While nucleoli appeared intact in Nopp140-/- cells, the C/D small nucleolar ribonucleoprotein (snoRNP) methyltransferase, fibrillarin, redistributed to the nucleoplasm in variable amounts depending on the cell type; RT-PCRs showed that 2'-O-methylation of ribosomal RNA (rRNA) in Nopp140-/- cells was reduced at select sites within both the 18S and 28S rRNAs. Ultrastructural analysis showed that Nopp140-/- cells were deficient in cytoplasmic ribosomes, but instead contained abnormal electron-dense cytoplasmic granules. Immunoblot analysis showed a loss of RpL34, and metabolic labeling showed a significant drop in protein translation, supporting the loss of functional ribosomes. Northern blots showed that pre-RNA cleavage pathways were generally unaffected by the loss of Nopp140, but that R2 retrotransposons that naturally reside within the 28S region of normally silent heterochromatic Drosophila ribosomal DNA (rDNA) genes were selectively expressed in Nopp140-/- larvae.…

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Crossref · 10.1007/s00412-014-0490-9

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Authors
He, F., James, A., Raje, H., Ghaffari, H., & DiMario, P.
Affiliations
Louisiana State University (US)
Corresponding author
DiMario, Patrick
Venue
Chromosoma
Published
2014-11-11
Volume
124
Issue
2
Pages
191-208
ISSN
  • 0009-5915 (ISSN-L)
  • 1432-0886
OpenAlex
W2027977110
Cited by
27
References
2
Citations by year
2026: 2 · 2025: 4 · 2024: 1 · 2023: 1 · 2022: 2 · 2021: 5 · 2020: 1 · 2019: 2 · 2018: 3 · 2017: 3 · 2016: 3
FWCI
0.82
Type
journal article
Language
en
Open access
closed
Locations
  • Chromosoma (publishedVersion)
  • PubMed (publishedVersion)
  • Civil War Book Review (submittedVersion)
Funders
National Science Foundation
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
Keywords
Nopp140, ribosomopathies, ribosome assembly, Drosophila, fibrillarin, nucleolus, R2 retrotransposon, mRNA translation, cytoplasmic ribosomes, developmental lethality
MeSH terms
Animals, Cell Nucleus, Chromatin, Chromosomal Proteins, Non-Histone, Drosophila, Female, Male, Nuclear Proteins, Phosphoproteins, Ribosomes, RNA, Ribosomal, Polymerase Chain Reaction, RNA-Binding Proteins, Gene Deletion, Coiled Bodies, Ribonucleoproteins, Small Nucleolar, Genomics, Drosophila Proteins

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