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https://www.readbyqxmd.com/read/29752352/functional-analysis-of-hsh155-sf3b1-interactions-with-the-u2-snrna-branch-site-duplex
#1
Tucker Joe Carrocci, Joshua Clark Paulson, Aaron Andrew Hoskins
SF3b1 is an essential component of the U2 snRNP implicated in branch site (BS) recognition and found to be frequently mutated in several human cancers. While recent structures of yeast and human SF3b1 have revealed its molecular architecture, the importance of specific RNA:protein contacts and conformational changes remain largely uncharacterized. Here, we performed mutational analysis of yeast SF3b1, guided by recent structures of the spliceosome. We find that conserved amino acids contacting the U2 snRNA backbone of the U2/BS duplex are nonessential, and that yeast can tolerate truncation of the HEAT repeats containing these amino acids...
May 11, 2018: RNA
https://www.readbyqxmd.com/read/29741121/when-sumo-met-splicing
#2
Berta Pozzi, Pablo Mammi, Laureano Bragado, Luciana E Giono, Anabella Srebrow
Spliceosomal proteins have been revealed as SUMO conjugation targets. Moreover, we have reported that many of these are in a SUMO-conjugated form when bound to a pre-mRNA substrate during a splicing reaction. We demonstrated that SUMOylation of Prp3 (PRPF3), a component of the U4/U6 di-snRNP, is required for U4/U6•U5 tri-snRNP formation and/or recruitment to active spliceosomes. Expanding upon our previous results, we have shown that the splicing factor SRSF1 stimulates SUMO conjugation to several spliceosomal proteins...
May 9, 2018: RNA Biology
https://www.readbyqxmd.com/read/29739806/the-surveillance-of-pre-mrna-splicing-is-an-early-step-in-c-elegans-rnai-of-endogenous-genes
#3
Martin A Newman, Fei Ji, Sylvia E J Fischer, Anthony Anselmo, Ruslan I Sadreyev, Gary Ruvkun
RNAi pathways detect and silence foreign nucleic acids such as viruses as well as endogenous genes in many species. The phylogenetic profile across eukaryotes of proteins that mediate key steps in RNAi is correlated with the profiles of multiple mRNA splicing proteins and with intron number, suggesting that RNAi may surveil mRNA splicing to detect the divergent or absent introns of viruses. Here we examine the role of mRNA splicing in Caenorhabditis elegans RNAi . We found that viable null mutations in U1 and U2 small nuclear ribonucleic protein (snRNP)-specific splicing factor genes cause defects in RNAi...
May 8, 2018: Genes & Development
https://www.readbyqxmd.com/read/29717126/architecture-of-the-u6-snrnp-reveals-specific-recognition-of-3-end-processed-u6-snrna
#4
Eric J Montemayor, Allison L Didychuk, Allyson D Yake, Gurnimrat K Sidhu, David A Brow, Samuel E Butcher
The spliceosome removes introns from precursor messenger RNA (pre-mRNA) to produce mature mRNA. Prior to catalysis, spliceosomes are assembled de novo onto pre-mRNA substrates. During this assembly process, U6 small nuclear RNA (snRNA) undergoes extensive structural remodeling. The early stages of this remodeling process are chaperoned by U6 snRNP proteins Prp24 and the Lsm2-8 heteroheptameric ring. We now report a structure of the U6 snRNP from Saccharomyces cerevisiae. The structure reveals protein-protein contacts that position Lsm2-8 in close proximity to the chaperone "active site" of Prp24...
May 1, 2018: Nature Communications
https://www.readbyqxmd.com/read/29715471/functional-and-biochemical-characterization-of-dib1-s-role-in-pre-messenger-rna-splicing
#5
Christian C Schreib, Emily K Bowman, Cody A Hernandez, Amber L Lucas, Camille H S Potts, Corina Maeder
The spliceosome is a dynamic macromolecular machine that undergoes a series of conformational rearrangements as it transitions between the several states required for accurate splicing. The transition from the B to Bact is a key part of spliceosome assembly and is defined by the departure of several proteins, including essential U5 component Dib1. Recent structural studies suggest that Dib1 has a role in preventing premature spliceosome activation, as it is positioned adjacent to the U6 snRNA ACAGAGA and the U5 loop I, but its mechanism is unknown...
April 28, 2018: Journal of Molecular Biology
https://www.readbyqxmd.com/read/29684050/intronic-pah-gene-mutations-cause-a-splicing-defect-by-a-novel-mechanism-involving-u1snrnp-binding-downstream-of-the-5-splice-site
#6
Ainhoa Martínez-Pizarro, Maja Dembic, Belén Pérez, Brage S Andresen, Lourdes R Desviat
Phenylketonuria (PKU), one of the most common inherited diseases of amino acid metabolism, is caused by mutations in the phenylalanine hydroxylase (PAH) gene. Recently, PAH exon 11 was identified as a vulnerable exon due to a weak 3' splice site, with different exonic mutations affecting exon 11 splicing through disruption of exonic splicing regulatory elements. In this study, we report a novel intron 11 regulatory element, which is involved in exon 11 splicing, as revealed by the investigated pathogenic effect of variants c...
April 23, 2018: PLoS Genetics
https://www.readbyqxmd.com/read/29671102/phosphorylation-by-prp4-kinase-releases-the-self-inhibition-of-fgprp31-in-fusarium-graminearum
#7
Xuli Gao, Ju Zhang, Chaoni Song, Kangyi Yuan, Jianhua Wang, Qiaojun Jin, Jin-Rong Xu
Prp31 is one of the key tri-snRNP components essential for pre-mRNA splicing although its exact molecular function is not well studied. In a previous study, suppressor mutations were identified in the PRP31 ortholog in two spontaneous suppressors of Fgprp4 mutant deleted of the only kinase of the spliceosome in Fusarium graminearum. To further characterize the function of FgPrp31 and its relationship with FgPrp4 kinase, in this study we identified additional suppressor mutations in FgPrp31 and determined the suppressive effects of selected mutations...
April 18, 2018: Current Genetics
https://www.readbyqxmd.com/read/29643343/author-correction-cryoem-structure-of-saccharomyces-cerevisiae-u1-snrnp-offers-insight-into-alternative-splicing
#8
Xueni Li, Shiheng Liu, Jiansen Jiang, Lingdi Zhang, Sara Espinosa, Ryan C Hill, Kirk C Hansen, Z Hong Zhou, Rui Zhao
The originally published version of this Article contained several errors in Figure 2, panel a: the basepair register in SL3-4 of yeast U1 snRNA was depicted incorrectly; the basepair for A287-U295 in yeast U1 snRNA was erroneously present; basepairs for U84-G119, G309-U532, A288-U295 and U289-A294 in yeast U1 snRNA were missing; the bulging nucleotide in SL3 of human U1 snRNA was depicted as G instead of C; and the dashed boxes defining the 5' ss binding site and Sm site in both human and yeast snRNAs were not drawn accurately...
April 11, 2018: Nature Communications
https://www.readbyqxmd.com/read/29628886/u1-snrnp-alteration-and-neuronal-cell-cycle-reentry-in-alzheimer-disease
#9
REVIEW
Bing Bai
The aberrancy of U1 small nuclear ribonucleoprotein (snRNP) complex and RNA splicing has been demonstrated in Alzheimer's disease (AD). Importantly, the U1 proteopathy is AD-specific, widespread and early-occurring, thus providing a very unique clue to the AD pathogenesis. The prominent feature of U1 histopathology is its nuclear depletion and redistribution in the neuronal cytoplasm. According to the preliminary data, the initial U1 cytoplasmic distribution pattern is similar to the subcellular translocation of the spliceosome in cells undergoing mitosis...
2018: Frontiers in Aging Neuroscience
https://www.readbyqxmd.com/read/29618593/a-genetic-screen-for-pre-mrna-splicing-mutants-of-arabidopsis-thaliana-identifies-putative-u1-snrnp-components-rbm25-and-prp39a
#10
(no author information available yet)
No abstract text is available yet for this article.
April 2018: Genetics
https://www.readbyqxmd.com/read/29615482/defining-essential-elements-and-genetic-interactions-of-the-yeast-lsm2-8-ring-and-demonstration-that-essentiality-of-lsm2-8-is-bypassed-via-overexpression-of-u6-snrna-or-the-u6-snrnp-subunit-prp24
#11
Allen J Roth, Stewart Shuman, Beate Schwer
A seven-subunit Lsm2-8 protein ring assembles on the U-rich 3' end of the U6 snRNA. A structure-guided mutational analysis of the Saccharomyces cerevisiae Lsm2-8 ring affords new insights to structure-function relations and genetic interactions of the Lsm subunits. Alanine scanning of 39 amino acids comprising the RNA-binding sites or inter-subunit interfaces of Lsm2, Lsm3, Lsm4, Lsm5, and Lsm8 identified only one instance of lethality (Lsm3-R69A) and one severe growth defect (Lsm2-R63A), both involving amino acids that bind the 3'-terminal UUU trinucleotide...
April 3, 2018: RNA
https://www.readbyqxmd.com/read/29529248/protein-composition-of-catalytically-active-u7-dependent-processing-complexes-assembled-on-histone-pre-mrna-containing-biotin-and-a-photo-cleavable-linker
#12
Aleksandra Skrajna, Xiao-Cui Yang, Michal Dadlez, William F Marzluff, Zbigniew Dominski
3' end cleavage of metazoan replication-dependent histone pre-mRNAs requires the multi-subunit holo-U7 snRNP and the stem-loop binding protein (SLBP). The exact composition of the U7 snRNP and details of SLBP function in processing remain unclear. To identify components of the U7 snRNP in an unbiased manner, we developed a novel approach for purifying processing complexes from Drosophila and mouse nuclear extracts. In this method, catalytically active processing complexes are assembled in vitro on a cleavage-resistant histone pre-mRNA containing biotin and a photo-sensitive linker, and eluted from streptavidin beads by UV irradiation for direct analysis by mass spectrometry...
February 26, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29507115/neurochondrin-interacts-with-the-smn-protein-suggesting-a-novel-mechanism-for-spinal-muscular-atrophy-pathology
#13
Luke W Thompson, Kim D Morrison, Sally L Shirran, Ewout J N Groen, Thomas H Gillingwater, Catherine H Botting, Judith E Sleeman
Spinal muscular atrophy (SMA) is an inherited neurodegenerative condition caused by a reduction in the amount of functional survival motor neuron (SMN) protein. SMN has been implicated in transport of mRNA in neural cells for local translation. We previously identified microtubule-dependent mobile vesicles rich in SMN and SNRPB, a member of the Sm family of small nuclear ribonucleoprotein (snRNP)-associated proteins, in neural cells. By comparing the interactomes of SNRPB and SNRPN, a neural-specific Sm protein, we now show that the essential neural protein neurochondrin (NCDN) interacts with Sm proteins and SMN in the context of mobile vesicles in neurites...
April 17, 2018: Journal of Cell Science
https://www.readbyqxmd.com/read/29491137/the-cryo-em-structure-of-the-sf3b-spliceosome-complex-bound-to-a-splicing-modulator-reveals-a-pre-mrna-substrate-competitive-mechanism-of-action
#14
Lorenzo I Finci, Xiaofeng Zhang, Xiuliang Huang, Qiang Zhou, Jennifer Tsai, Teng Teng, Anant Agrawal, Betty Chan, Sean Irwin, Craig Karr, Andrew Cook, Ping Zhu, Dominic Reynolds, Peter G Smith, Peter Fekkes, Silvia Buonamici, Nicholas A Larsen
Somatic mutations in spliceosome proteins lead to dysregulated RNA splicing and are observed in a variety of cancers. These genetic aberrations may offer a potential intervention point for targeted therapeutics. SF3B1, part of the U2 small nuclear RNP (snRNP), is targeted by splicing modulators, including E7107, the first to enter clinical trials, and, more recently, H3B-8800. Modulating splicing represents a first-in-class opportunity in drug discovery, and elucidating the structural basis for the mode of action opens up new possibilities for structure-based drug design...
February 1, 2018: Genes & Development
https://www.readbyqxmd.com/read/29425494/crosstalk-between-the-rna-methylation-and-histone-binding-activities-of-mepce-regulates-p-tefb-activation-on-chromatin
#15
Samantha B Shelton, Nakul M Shah, Nathan S Abell, Sravan K Devanathan, Marvin Mercado, Blerta Xhemalçe
RNAP II switching from the paused to the productive transcription elongation state is a pivotal regulatory step that requires specific phosphorylations catalyzed by the P-TEFb kinase. Nucleosolic P-TEFb activity is inhibited by its interaction with the ribonuclear protein complex built around the 7SK small nuclear RNA (7SK snRNP). MePCE is the RNA methyltransferase that methylates and stabilizes 7SK in the nucleosol. Here, we report that MePCE also binds chromatin through the histone H4 tail to serve as a P-TEFb activator at specific genes important for cellular identity...
February 6, 2018: Cell Reports
https://www.readbyqxmd.com/read/29415178/the-sm-core-mediates-the-retention-of-partially-assembled-spliceosomal-snrnps-in-cajal-bodies-until-their-full-maturation
#16
Adriana Roithová, Klára Klimešová, Josef Pánek, Cindy L Will, Reinhard Lührmann, David Stanek, Cyrille Girard
Cajal bodies (CBs) are nuclear non-membrane bound organelles where small nuclear ribonucleoprotein particles (snRNPs) undergo their final maturation and quality control before they are released to the nucleoplasm. However, the molecular mechanism how immature snRNPs are targeted and retained in CBs has yet to be described. Here, we microinjected and expressed various snRNA deletion mutants as well as chimeric 7SK, Alu or bacterial SRP non-coding RNAs and provide evidence that Sm and SMN binding sites are necessary and sufficient for CB localization of snRNAs...
February 5, 2018: Nucleic Acids Research
https://www.readbyqxmd.com/read/29398447/transcriptional-pause-sites-delineate-stable-nucleosome-associated-premature-polyadenylation-suppressed-by-u1-snrnp
#17
Anthony C Chiu, Hiroshi I Suzuki, Xuebing Wu, Dig B Mahat, Andrea J Kriz, Phillip A Sharp
Regulation of RNA polymerase II (Pol II) elongation is a critical step in gene regulation. Here, we report that U1 snRNP recognition and transcription pausing at stable nucleosomes are linked through premature polyadenylation signal (PAS) termination. By generating RNA exosome conditional deletion mouse embryonic stem cells, we identified a large class of polyadenylated short transcripts in the sense direction destabilized by the RNA exosome. These PAS termination events are enriched at the first few stable nucleosomes flanking CpG islands and suppressed by U1 snRNP...
February 15, 2018: Molecular Cell
https://www.readbyqxmd.com/read/29381447/stabilized1-as-a-heat-stress-specific-splicing-factor-in-arabidopsis-thaliana
#18
Geun-Don Kim, Sang-Dong Yoo, Young-Hee Cho
To overcome high temperature stress, plants have developed transcriptional cascades which express a large amount of chaperone proteins called heat shock proteins (HSPs). In our recent publication, we reported that STABILIZED1, as an U5-snRNP-interacting protein, is involved in the splicing of heat shock factor (HSF) and HSP transcripts during high temperature stress. This indicates that not only transcriptional regulation, but also post-transcriptional regulation by STA1, is essential for the full activation of HSF-HSP cascades and for thermotolerance...
February 1, 2018: Plant Signaling & Behavior
https://www.readbyqxmd.com/read/29371219/-gemin4-is-an-essential-gene-in-mice-and-its-overexpression-in-human-cells-causes-relocalization-of-the-smn-complex-to-the-nucleoplasm
#19
Ingo D Meier, Michael P Walker, A Gregory Matera
Gemin4 is a member of the Survival Motor Neuron (SMN) protein complex, which is responsible for the assembly and maturation of Sm-class small nuclear ribonucleoproteins (snRNPs). In metazoa, Sm snRNPs are assembled in the cytoplasm and subsequently imported into the nucleus. We previously showed that the SMN complex is required for snRNP import in vitro , although it remains unclear which specific components direct this process. Here, we report that Gemin4 overexpression drives SMN and the other Gemin proteins from the cytoplasm into the nucleus...
February 1, 2018: Biology Open
https://www.readbyqxmd.com/read/29360106/structure-of-the-human-activated-spliceosome-in-three-conformational-states
#20
Xiaofeng Zhang, Chuangye Yan, Xiechao Zhan, Lijia Li, Jianlin Lei, Yigong Shi
During each cycle of pre-mRNA splicing, the pre-catalytic spliceosome (B complex) is converted into the activated spliceosome (Bact complex), which has a well-formed active site but cannot proceed to the branching reaction. Here, we present the cryo-EM structure of the human Bact complex in three distinct conformational states. The EM map allows atomic modeling of nearly all protein components of the U2 small nuclear ribonucleoprotein (snRNP), including three of the SF3a complex and seven of the SF3b complex...
March 2018: Cell Research
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