A5012957405- RNA Research and Splicing
- RNA modifications and cancer
- RNA and protein synthesis mechanisms
- RNA regulation and disease
- Neurogenetic and Muscular Disorders Research
- Nuclear Structure and Function
- Genomics and Chromatin Dynamics
- RNA Interference and Gene Delivery
- MicroRNA in disease regulation
- Cancer-related molecular mechanisms research
- Advanced biosensing and bioanalysis techniques
- CRISPR and Genetic Engineering
- DNA Repair Mechanisms
- Chromosomal and Genetic Variations
- Retinal Development and Disorders
- Diagnosis and treatment of tuberculosis
- Biochemical and Molecular Research
- Immune Cell Function and Interaction
- Renal and related cancers
- Immunodeficiency and Autoimmune Disorders
- Microtubule and mitosis dynamics
- Trypanosoma species research and implications
- interferon and immune responses
- ATP Synthase and ATPases Research
- Protein Degradation and Inhibitors
Czech Academy of Sciences
2012-2025
Czech Academy of Sciences, Institute of Molecular Genetics
2015-2025
Institute of Molecular Genetics
2008-2023
O2 Czech Republic (Czechia)
2023
Czech Academy of Sciences, J. Heyrovský Institute of Physical Chemistry
2010
Charles University
1997-2008
Max Planck Institute of Molecular Cell Biology and Genetics
2003-2006
Czech Academy of Sciences, Institute of Physiology
2006
Max Planck Society
2005
Czech Academy of Sciences, Institute of Experimental Medicine
1998-2001
Cajal bodies (CBs) are subnuclear domains implicated in small nuclear ribonucleoprotein (snRNP) biogenesis. In most cell types, CBs coincide with gems, which contain the survival of motor neurons (SMN) complex, an essential snRNP assembly factor. Here, we analyze exchange kinetics multiple components and gems living cells using photobleaching microscopy. We demonstrate differences dissociation CB constituents relate them to their functions. Coilin SMN complex members exhibit relatively long...
There is increasing evidence to suggest that splicing decisions are largely made when the nascent RNA still associated with chromatin. Here we demonstrate activity of histone deacetylases (HDACs) influences splice site selection. Using splicing-sensitive microarrays, identified ∼700 genes whose was altered after HDAC inhibition. We provided inhibition induced H4 acetylation and increased Polymerase II (Pol II) processivity along an alternatively spliced element. In addition, reduced...
Precursor messenger RNA (pre-mRNA) splicing is catalyzed by the spliceosome, a large ribonucleoprotein (RNP) complex composed of five small nuclear RNP particles (snRNPs) and additional proteins. Using live cell imaging GFP-tagged snRNP components expressed at endogenous levels, we examined how spliceosome assembles in vivo. A comprehensive analysis dynamics nucleus enabled us to determine diffusion throughout nucleoplasm as well interaction rates individual snRNPs with pre-mRNA. Core U2 U5...
Splicing is catalyzed by the spliceosome, a complex of five major small nuclear ribonucleoprotein particles (snRNPs). The pre-mRNA splicing factor PRPF8 crucial component U5 snRNP, and together with EFTUD2 SNRNP200, it forms central module spliceosome. Using quantitative proteomics, we identified assembly intermediates containing PRPF8, EFTUD2, SNRNP200 in association HSP90/R2TP complex, its ZNHIT2 cofactor, additional proteins. HSP90 R2TP bind unassembled proteins cytoplasm, stabilize them,...
Many nascent long non-coding RNAs (lncRNAs) undergo the same maturation steps as pre-mRNAs of protein-coding genes (PCGs), but they are often poorly spliced. To identify underlying mechanisms for this phenomenon, we searched putative splicing inhibitory sequences using ncRNA-a2 a model. Genome-wide analyses intergenic lncRNAs (lincRNAs) revealed that lincRNA efficiency positively correlates with 5′ss strength while no such correlation was identified PCGs. In addition, efficiently spliced...
The spliceosomal small nuclear RNAs (snRNAs) are distributed throughout the nucleoplasm and concentrated in inclusions termed Cajal bodies (CBs). A role for CBs metabolism of snRNPs has been proposed but is not well understood. SART3/p110 protein interacts transiently with U6 U4/U6 promotes reassembly after splicing vitro. Here we report that enriched gems or residual lacking coilin. snRNP Sm-like (LSm) proteins, also involved assembly, were localized to as well. levels LSm proteins reduced...
Spliceosomal small nuclear ribonucleoprotein particles (snRNPs) are required for pre-mRNA splicing throughout the nucleoplasm, yet snRNPs also concentrate in Cajal bodies (CBs). To address a proposed role of CBs snRNP assembly, we have used fluorescence resonance energy transfer (FRET) microscopy to investigate subnuclear distribution specific intermediates. Two distinct complexes containing protein SART3 (p110), U4/U6 were localized: SART3•U6 and SART3•U4/U6 snRNP. These segregated...
Spliceosomal small nuclear ribonucleoprotein particles (snRNPs) undergo specific assembly steps in Cajal bodies (CBs), nonmembrane-bound compartments within cell nuclei. An example is the U4/U6 di-snRNP, assembled from U4 and U6 monomers. These snRNPs can also assemble nucleoplasm when cells lack CBs. Here, we address hypothesis that snRNP concentration CBs facilitates assembly, by comparing predicted rates of association nuclei with without This was accomplished a random walk-and-capture...
The Cajal body (CB) is a nuclear structure closely associated with import and biogenesis of small ribonucleoprotein particles (snRNPs). Here, we tested whether CBs also contain mature snRNPs CB integrity depends on the ongoing snRNP splicing cycle. Sm proteins tagged photoactivatable color-maturing variants fluorescent were used to monitor behavior in living cells over time; accumulated CBs, traveled from one another, they not preferentially replaced by newly imported snRNPs. To test cycle,...
Cajal bodies (CBs) are evolutionarily conserved nuclear structures involved in the metabolism of spliceosomal small ribonucleoprotein particles (snRNPs). CBs not present all cell types, and trigger for their formation is yet known. Here, we depleted cells factors required final steps snRNP assembly assayed presence stalled intermediates CBs. We show that depletion induces normally lack these compartments, suggesting CB nucleation triggered by an imbalance assembly. Accumulation depends on...
Brd2 is a member of the bromodomain extra terminal (BET) protein family, which consists four chromatin-interacting proteins that regulate gene expression. Each BET contains two N-terminal bromodomains, recognize acetylated histones, and C-terminal protein–protein interaction domain. Using genome-wide screen, we identify 1450 genes whose transcription regulated by Brd2. In addition, almost 290 change their alternative splicing pattern upon depletion. specifically localized at promoters target...
Abstract Spliceosomal snRNPs are multicomponent particles that undergo a complex maturation pathway. Human Sm-class snRNAs generated as 3′-end extended precursors, which exported to the cytoplasm and assembled together with Sm proteins into core RNPs by SMN complex. Here, we provide evidence these pre-snRNA substrates contain compact, evolutionarily conserved secondary structures overlap binding site. These structural motifs in pre-snRNAs predicted interfere assembly. We model rearrangements...
Retinitis pigmentosa (RP) is a hereditary disorder caused by mutations in more than 70 different genes including those that encode proteins important for pre-mRNA splicing. Most RP-associated splicing factors reduce either their expression, stability or incorporation into functional complexes. However, we have previously shown two RP PRPF8 (F2314L and Y2334N) SNRNP200 (S1087L R1090L) behaved differently, it was still unclear how these affect the functions of both proteins. To investigate...
PUF60 is a splicing factor that binds uridine (U)-rich tracts and facilitates association of the U2 small nuclear ribonucleoprotein with primary transcripts. deficiency (PD) causes developmental delay coupled intellectual disability spinal, cardiac, ocular renal defects, but PD pathogenesis not understood. Using RNA-Seq, we identify human PUF60-regulated exons show preferentially acts as their activator. PUF60-activated internal are enriched for Us upstream 3′ splice sites (3′ss), preceded...
Abstract Mutations in BRAT1 , encoding BRCA1-associated ATM activator 1, have been associated with neurodevelopmental and neurodegenerative disorders characterized by heterogeneous phenotypes varying levels of clinical severity. However, the underlying molecular mechanisms disease pathology remain poorly understood. Here, we show that tightly interacts INTS9/INTS11 subunits Integrator complex processes 3’ ends various noncoding RNAs pre-mRNAs. We find functions are disrupted deletion. In...