scholarly journals Loss of U11 small nuclear RNA in the developing mouse limb results in micromelia

Development ◽  
2020 ◽  
Vol 147 (21) ◽  
pp. dev190967
Author(s):  
Kyle D. Drake ◽  
Christopher Lemoine ◽  
Gabriela S. Aquino ◽  
Anna M. Vaeth ◽  
Rahul N. Kanadia
Keyword(s):  
Cell Cycle ◽  
Limb Development ◽  
Intron Retention ◽  
Size Control ◽  
Small Nuclear Rna ◽  
Essential Components ◽  
Primordial Dwarfism ◽  
Nuclear Rna ◽  
Mouse Limb

ABSTRACTDisruption of the minor spliceosome due to mutations in RNU4ATAC is linked to primordial dwarfism in microcephalic osteodysplastic primordial dwarfism type 1, Roifman syndrome, and Lowry-Wood syndrome. Similarly, primordial dwarfism in domesticated animals is linked to positive selection in minor spliceosome components. Despite being vital for limb development and size regulation, its role remains unexplored. Here, we disrupt minor spliceosome function in the developing mouse limb by ablating one of its essential components, U11 small nuclear RNA, which resulted in micromelia. Notably, earlier loss of U11 corresponded to increased severity. We find that limb size is reduced owing to elevated minor intron retention in minor intron-containing genes that regulate cell cycle. As a result, limb progenitor cells experience delayed prometaphase-to-metaphase transition and prolonged S-phase. Moreover, we observed death of rapidly dividing, distally located progenitors. Despite cell cycle defects and cell death, the spatial expression of key limb patterning genes was maintained. Overall, we show that the minor spliceosome is required for limb development via size control potentially shared in disease and domestication.

scholarly journals Minor spliceosome disruption causes limb growth defects without altering patterning

2020 ◽  
Author(s):  
Kyle D Drake ◽  
Christopher Lemoine ◽  
Gabriela S Aquino ◽  
Anna M Vaeth ◽  
Rahul N Kanadia
Keyword(s):  
Limb Development ◽  
Intron Retention ◽  
S Phase ◽  
Small Nuclear Rna ◽  
Potential Mechanism ◽  
Limb Patterning ◽  
Growth Defects ◽  
Primordial Dwarfism ◽  
Nuclear Rna

AbstractDisruption of the minor spliceosome causes primordial dwarfism in microcephalic osteodysplastic primordial dwarfism type 1. Similarly, primordial dwarfism in domesticated animals is linked to positive selection in minor spliceosome components. Despite the importance of minor intron splicing in limb size regulation, its role in limb development remains unexplored. Here we show that loss of U11 small nuclear RNA, an essential minor spliceosome component, results in stunted limbs that maintain patterning. Notably, earlier loss of U11 corresponded to increased severity. We find that limb size is reduced due to elevated minor intron retention in minor intron-containing genes that regulate cell cycle. Limb progenitor cells experience delayed prometaphase to metaphase transition and prolonged S-phase, resulting in death of rapidly dividing, distally located progenitors. Consequently, crucial limb patterning genes are upregulated and their expression is maintained spatially to achieve basic patterning. Overall, these findings reveal a potential mechanism shared in disease and domestication.

scholarly journals Trp53 ablation fails to prevent microcephaly in mouse pallium with impaired minor intron splicing

2021 ◽  
Author(s):  
Alisa K. White ◽  
Marybeth Baumgartner ◽  
Madisen F. Lee ◽  
Kyle D. Drake ◽  
Gabriela S. Aquino ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Damage ◽  
Intron Retention ◽  
Conditional Knockout ◽  
Intron Splicing ◽  
Small Nuclear Rna ◽  
Primary Microcephaly ◽  
Double Knockout ◽  
Nuclear Rna

AbstractMutations in minor spliceosome component RNU4ATAC, a small nuclear RNA (snRNA), are linked to primary microcephaly. We have reported that in the conditional knockout (cKO) mice for Rnu11, another minor spliceosome snRNA, minor intron splicing defect in minor intron-containing genes (MIGs) regulating cell cycle resulted in cell cycle defects, with a concomitant increase in γH2aX+ cells and p53-mediated apoptosis. Trp53 ablation in the Rnu11 cKO mice did not prevent microcephaly. However, RNAseq analysis of the double knockout (dKO) pallium reflected transcriptomic shift towards the control from the Rnu11 cKO. We found elevated minor intron retention and alternative splicing across minor introns in the dKO. Disruption of these MIGs resulted in cell cycle defects that were more severe and detected earlier in the dKO, but with delayed detection of γH2aX+ DNA damage. Thus, p53 might also play a role in causing DNA damage in the developing pallium. In all, our findings further refine our understanding of the role of the minor spliceosome in cortical development and identify MIGs underpinning microcephaly in minor spliceosome-related diseases.

scholarly journals Minor spliceosome inactivation in the developing mouse cortex causes self-amplifying radial glial cell death and microcephaly

2017 ◽  
Author(s):  
Marybeth Baumgartner ◽  
Anouk M. Olthof ◽  
Katery C. Hyatt ◽  
Christopher Lemoine ◽  
Kyle Drake ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Glial Cell ◽  
Progenitor Cell ◽  
Intron Retention ◽  
Cortical Development ◽  
Conditional Knockout ◽  
Small Nuclear Rna ◽  
Radial Glial Cell ◽  
Nuclear Rna ◽  
Radial Glial

AbstractInactivation of the minor spliceosome has been linked to microcephalic osteodysplastic primordial dwarfism type 1 (MOPD1). To interrogate how minor intron splicing regulates cortical development, we employed Emx1-Cre to ablate Rnu11, which encodes the minor spliceosome-specific U11 small nuclear RNA (snRNA), in the developing cortex (pallium). Rnu11 cKO mice were born with microcephaly, caused by death of self-amplifying radial glial cells (RGCs). However, both intermediate progenitor cells (IPCs) and neurons were produced in the U11-null pallium. RNAseq of the pallium revealed elevated minor intron retention in the mutant, particularly in genes regulating cell cycle. Moreover, the only downregulated minor intron-containing gene (MIG) was Spc24, which regulates kinetochore assembly. These findings were consistent with the observation of fewer RGCs entering cytokinesis prior to RGC loss, underscoring the requirement of minor splicing for cell cycle progression in RGCs. Overall, we provide a potential explanation of how disruption of minor splicing might cause microcephaly in MOPD1.Summary StatementHere we report the first mammalian model to investigate the role of the minor spliceosome in cortical development and microcephaly.List of abbreviations usedMOPD1=microcephalic osteodysplastic primordial dwarfism type 1; snRNA=small nuclear RNA; cKO=conditional knockout; NPC=neural progenitor cell; RGC=radial glial cell; IPC=intermediate progenitor cell; MIG=minor intron-containing gene

Cell cycle redistribution of U3 snRNA and fibrillarin. Presence in the cytoplasmic nucleolus remnant and in the prenucleolar bodies at telophase

1994 ◽  
Vol 107 (2) ◽  
pp. 463-475 ◽  
Author(s):  
M.C. Azum-Gelade ◽  
J. Noaillac-Depeyre ◽  
M. Caizergues-Ferrer ◽  
N. Gas
Keyword(s):  
Cell Cycle ◽  
Immunofluorescence Microscopy ◽  
Close Association ◽  
Ultrastructural Level ◽  
Small Nuclear Rna ◽  
Cho Cell ◽  
Nucleolar Proteins ◽  
Nuclear Rna ◽  
Active Nors

The distribution of the U3 small nuclear RNA during the cell cycle of the CHO cell line was studied by in situ hybridization using digoxigenin-labelled oligonucleotide probes. The location of the hybrids by immunofluorescence microscopy and at the ultrastructural level was correlated with the distribution of two nucleolar proteins, nucleolin and fibrillarin. The U3 snRNA molecules persist throughout mitosis in close association with the nucleolar remnant. U3 snRNA is present in the prenucleolar bodies (PNBs) and could participate in nucleologenesis in association with several nucleolar proteins such as nucleolin and fibrillarin. The interaction of U3 snRNP with the 5′ external spacer of pre-RNA newly synthesized by active NORs is proposed to be the promoting event of nucleologenesis.

scholarly journals RN7SK small nuclear RNA controls bidirectional transcription of highly expressed gene pairs in skin

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Roberto Bandiera ◽  
Rebecca E. Wagner ◽  
Thiago Britto-Borges ◽  
Christoph Dieterich ◽  
Sabine Dietmann ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Rna Polymerase Ii ◽  
Noncoding Rna ◽  
Stem Cell Differentiation ◽  
Cell Cycle Regulators ◽  
Small Nuclear Rna ◽  
Pol Ii ◽  
Gene Pairs ◽  
Nuclear Rna ◽  
Numerous Cell

AbstractPausing of RNA polymerase II (Pol II) close to promoters is a common regulatory step in RNA synthesis, and is coordinated by a ribonucleoprotein complex scaffolded by the noncoding RNA RN7SK. The function of RN7SK-regulated gene transcription in adult tissue homoeostasis is currently unknown. Here, we deplete RN7SK during mouse and human epidermal stem cell differentiation. Unexpectedly, loss of this small nuclear RNA specifically reduces transcription of numerous cell cycle regulators leading to cell cycle exit and differentiation. Mechanistically, we show that RN7SK is required for efficient transcription of highly expressed gene pairs with bidirectional promoters, which in the epidermis co-regulated cell cycle and chromosome organization. The reduction in transcription involves impaired splicing and RNA decay, but occurs in the absence of chromatin remodelling at promoters and putative enhancers. Thus, RN7SK is directly required for efficient Pol II transcription of highly transcribed bidirectional gene pairs, and thereby exerts tissue-specific functions, such as maintaining a cycling cell population in the epidermis.

scholarly journals Nuclear Domains Enriched in RNA 3′-processing Factors Associate with Coiled Bodies and Histone Genes in a Cell Cycle–dependent Manner

1999 ◽  
Vol 10 (11) ◽  
pp. 3815-3824 ◽  
Author(s):  
Wouter Schul ◽  
Ineke van der Kraan ◽  
A. Gregory Matera ◽  
Roel van Driel ◽  
Luitzen de Jong
Keyword(s):  
Cell Cycle ◽  
Gene Clusters ◽  
Dependent Manner ◽  
Histone Genes ◽  
Small Nuclear Rna ◽  
Coiled Bodies ◽  
Nuclear Rna ◽  
Cycle Dependent ◽  
Nuclear Domains ◽  
Processing Factors

Nuclear domains, called cleavage bodies, are enriched in the RNA 3′-processing factors CstF 64 kDa and and CPSF 100 kDa. Cleavage bodies have been found either overlapping with or adjacent to coiled bodies. To determine whether the spatial relationship between cleavage bodies and coiled bodies was influenced by the cell cycle, we performed cell synchronization studies. We found that in G1 phase cleavage bodies and coiled bodies were predominantly coincident, whereas in S phase they were mostly adjacent to each other. In G2 cleavage bodies were often less defined or absent, suggesting that they disassemble at this point in the cell cycle. A small number of genetic loci have been reported to be juxtaposed to coiled bodies, including the genes for U1 and U2 small nuclear RNA as well as the two major histone gene clusters. Here we show that cleavage bodies do not overlap with small nuclear RNA genes but do colocalize with the histone genes next to coiled bodies. These findings demonstrate that the association of cleavage bodies and coiled bodies is both dynamic and tightly regulated and suggest that the interaction between these nuclear neighbors is related to the cell cycle–dependent expression of histone genes.

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