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 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 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.

A fragile site in the human U2 small nuclear RNA gene cluster is revealed by adenovirus type 12 infection

1988 ◽  
Vol 8 (5) ◽  
pp. 1863-1867
Author(s):  
D M Durnam ◽  
J C Menninger ◽  
S H Chandler ◽  
P P Smith ◽  
J K McDougall
Keyword(s):  
Gene Cluster ◽  
Fragile Site ◽  
Maximum Size ◽  
Adenovirus Type ◽  
Fragile Sites ◽  
Similar Data ◽  
Small Nuclear Rna ◽  
Structural Alterations ◽  
Nuclear Rna

Using in situ hybridization, we found that the U2 small nuclear RNA gene cluster mapped very close to and was frequently disrupted by the gaps and breaks induced specifically in the human 17q21-q22 region by highly oncogenic adenovirus type 12 (Ad12). Restriction mapping revealed no structural alterations in the U2 gene locus as a result of Ad12 infection. Likewise, no Ad12-induced alterations in U2 RNA levels were detected. We estimate that the maximum size of the region specifically disrupted by this virus was less than 350 to 700 kilobases. A comparison of these data with similar data regarding biochemically induced fragile sites was made.

Localization of human U1 small nuclear RNA genes to band p36.3 of chromosome 1 by in situ hybridization

1984 ◽  
Vol 10 (3) ◽  
pp. 307-313 ◽  
Author(s):  
S. L. Naylor ◽  
B. U. Zabel ◽  
T. Manser ◽  
R. Gesteland ◽  
A. Y. Sakaguchi
Keyword(s):  
In Situ Hybridization ◽  
Chromosome 1 ◽  
Small Nuclear Rna ◽  
Nuclear Rna ◽  
Rna Genes

scholarly journals Human U1 small nuclear RNA pseudogenes do not map to the site of the U1 genes in 1p36 but are clustered in 1q12-q22.

1985 ◽  
Vol 5 (9) ◽  
pp. 2172-2180 ◽  
Author(s):  
V Lindgren ◽  
L B Bernstein ◽  
A M Weiner ◽  
U Francke
Keyword(s):  
In Situ Hybridization ◽  
Class I ◽  
Chromosome 1 ◽  
Small Nuclear Rna ◽  
Coding Region ◽  
Cell Biol ◽  
Gene Copies ◽  
Nuclear Rna ◽  
Human Chromosome 1

Human U1 small nuclear RNA is encoded by approximately 30 gene copies. All of the U1 genes share several kilobases of essentially perfect flanking homology both upstream and downstream from the U1 coding region, but remarkably, for many U1 genes excellent flanking homology extends at least 24 kilobases upstream and 20 kilobases downstream. Class I U1 RNA pseudogenes are abundant in the human genome. These pseudogenes contain a complete but imperfect U1 coding region and possess extensive flanking homology to the true U1 genes. We mapped four class I pseudogenes by in situ hybridization to the long arm of chromosome 1, bands q12-q22, a region distinct from the site on the distal short arm of chromosome 1 to which the U1 genes have been previously mapped (Lund et al., Mol. Cell. Biol. 3:2211-2220, 1983; Naylor et al., Somat. Cell Mol. Genet. 10:307-313, 1984). We confirmed our in situ hybridization results by genomic blotting experiments with somatic cell hybrid lines with translocation products of human chromosome 1. These experiments provide further evidence that class I U1 pseudogenes and the true U1 genes are not interspersed. The results, along with those published elsewhere (Bernstein et al., Mol. Cell. Biol. 5:2159-2171, 1985), suggest that gene amplification may be responsible for the sequence homogeneity of the human U1 gene family.

scholarly journals A fragile site in the human U2 small nuclear RNA gene cluster is revealed by adenovirus type 12 infection.

1988 ◽  
Vol 8 (5) ◽  
pp. 1863-1867 ◽  
Author(s):  
D M Durnam ◽  
J C Menninger ◽  
S H Chandler ◽  
P P Smith ◽  
J K McDougall
Keyword(s):  
Gene Cluster ◽  
Fragile Site ◽  
Maximum Size ◽  
Adenovirus Type ◽  
Fragile Sites ◽  
Similar Data ◽  
Small Nuclear Rna ◽  
Structural Alterations ◽  
Nuclear Rna

Using in situ hybridization, we found that the U2 small nuclear RNA gene cluster mapped very close to and was frequently disrupted by the gaps and breaks induced specifically in the human 17q21-q22 region by highly oncogenic adenovirus type 12 (Ad12). Restriction mapping revealed no structural alterations in the U2 gene locus as a result of Ad12 infection. Likewise, no Ad12-induced alterations in U2 RNA levels were detected. We estimate that the maximum size of the region specifically disrupted by this virus was less than 350 to 700 kilobases. A comparison of these data with similar data regarding biochemically induced fragile sites was made.

Human U1 small nuclear RNA pseudogenes do not map to the site of the U1 genes in 1p36 but are clustered in 1q12-q22

1985 ◽  
Vol 5 (9) ◽  
pp. 2172-2180 ◽  
Author(s):  
V Lindgren ◽  
L B Bernstein ◽  
A M Weiner ◽  
U Francke
Keyword(s):  
In Situ Hybridization ◽  
Class I ◽  
Chromosome 1 ◽  
Small Nuclear Rna ◽  
Coding Region ◽  
Cell Biol ◽  
Gene Copies ◽  
Nuclear Rna ◽  
Human Chromosome 1

Human U1 small nuclear RNA is encoded by approximately 30 gene copies. All of the U1 genes share several kilobases of essentially perfect flanking homology both upstream and downstream from the U1 coding region, but remarkably, for many U1 genes excellent flanking homology extends at least 24 kilobases upstream and 20 kilobases downstream. Class I U1 RNA pseudogenes are abundant in the human genome. These pseudogenes contain a complete but imperfect U1 coding region and possess extensive flanking homology to the true U1 genes. We mapped four class I pseudogenes by in situ hybridization to the long arm of chromosome 1, bands q12-q22, a region distinct from the site on the distal short arm of chromosome 1 to which the U1 genes have been previously mapped (Lund et al., Mol. Cell. Biol. 3:2211-2220, 1983; Naylor et al., Somat. Cell Mol. Genet. 10:307-313, 1984). We confirmed our in situ hybridization results by genomic blotting experiments with somatic cell hybrid lines with translocation products of human chromosome 1. These experiments provide further evidence that class I U1 pseudogenes and the true U1 genes are not interspersed. The results, along with those published elsewhere (Bernstein et al., Mol. Cell. Biol. 5:2159-2171, 1985), suggest that gene amplification may be responsible for the sequence homogeneity of the human U1 gene family.

RNAs radiate from gene to cytoplasm as revealed by fluorescence in situ hybridization

1995 ◽  
Vol 108 (7) ◽  
pp. 2565-2572 ◽  
Author(s):  
R.W. Dirks ◽  
K.C. Daniel ◽  
A.K. Raap
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Distribution Patterns ◽  
Transcription Unit ◽  
Similar Distribution ◽  
Small Nuclear Rna ◽  
Barr Virus ◽  
Nuclear Rna ◽  
Nuclear Signal

Genes for Epstein-Barr virus, human cytomegalovirus immediate early antigen and luciferase are abundantly transcribed in Namalwa, rat 9G and X1 cells, respectively. The EBV transcripts and HCMV-IE transcripts are extensively spliced, while in the luciferase transcript only a small intron sequence has to be spliced out. EBV transcripts are hardly localized in the cytoplasm while the luciferase and HCMV-IE transcripts are present in the cytoplasm and translated into proteins. We have correlated these characteristics with nuclear RNA distribution patterns as seen by fluorescence in situ hybridization. Transcripts of the HCMV-IE transcription unit were shown to be present in a main nuclear signal in the form of a track or elongated dot and as small nuclear RNA signals that radiate from this site towards the cytoplasm. A similar distribution pattern of small RNA signals was observed for transcripts of the luciferase gene, whereas the main nuclear signal was always observed as a dot and never as a track or elongated dot. In Namalwa cells, EBV transcripts were only present as track-like signals. The results suggest that when the extent for splicing is high, unspliced or partially spliced mRNAs begin to occupy elongated dot or track-like domains in the vicinity of the gene. When the extent of splicing is low, splicing is completed co-transcriptionally, leading to a bright dot-like signal. The presence of small nuclear spots in addition to the main signal correlates with cytoplasmic mRNA expression. The small spots most likely represent, therefore, mRNAs in transport to the cytoplasm.

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 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

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