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[PMID]:29295980
[Au] Autor:Barragán-Iglesias P; Lou TF; Bhat VD; Megat S; Burton MD; Price TJ; Campbell ZT
[Ad] Endereço:School of Behavioral and Brain Sciences, University of Texas at Dallas, Richardson, TX, 75080, USA.
[Ti] Título:Inhibition of Poly(A)-binding protein with a synthetic RNA mimic reduces pain sensitization in mice.
[So] Source:Nat Commun;9(1):10, 2018 01 02.
[Is] ISSN:2041-1723
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Nociceptors rely on cap-dependent translation to rapidly induce protein synthesis in response to pro-inflammatory signals. Comparatively little is known regarding the role of the regulatory factors bound to the 3' end of mRNA in nociceptor sensitization. Poly(A)-binding protein (PABP) stimulates translation initiation by bridging the Poly(A) tail to the eukaryotic initiation factor 4F complex associated with the mRNA cap. Here, we use unbiased assessment of PABP binding specificity to generate a chemically modified RNA-based competitive inhibitor of PABP. The resulting RNA mimic, which we designated as the Poly(A) SPOT-ON, is more stable than unmodified RNA and binds PABP with high affinity and selectivity in vitro. We show that injection of the Poly(A) SPOT-ON at the site of an injury can attenuate behavioral response to pain. Collectively, these results suggest that PABP is integral for nociceptive plasticity. The general strategy described here provides a broad new source of mechanism-based inhibitors for RNA-binding proteins and is applicable for in vivo studies.
[Mh] Termos MeSH primário: Dor/metabolismo
Poli A/metabolismo
Proteínas de Ligação a Poli(A)/metabolismo
RNA/metabolismo
[Mh] Termos MeSH secundário: Animais
Linhagem Celular Tumoral
Células Cultivadas
Gânglios Espinais/citologia
Seres Humanos
Camundongos
Neurônios/efeitos dos fármacos
Neurônios/metabolismo
Dor Nociceptiva/metabolismo
Dor Nociceptiva/prevenção & controle
Dor/prevenção & controle
Medição da Dor
Poli A/química
Poli A/farmacologia
Proteínas de Ligação a Poli(A)/química
Ligação Proteica
RNA/química
RNA/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Poly(A)-Binding Proteins); 24937-83-5 (Poly A); 63231-63-0 (RNA)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180305
[Lr] Data última revisão:
180305
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180104
[St] Status:MEDLINE
[do] DOI:10.1038/s41467-017-02449-5


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[PMID]:28456523
[Au] Autor:Chikne V; Gupta SK; Doniger T; K SR; Cohen-Chalamish S; Waldman Ben-Asher H; Kolet L; Yahia NH; Unger R; Ullu E; Kolev NG; Tschudi C; Michaeli S
[Ad] Endereço:The Mina and Everard Goodman Faculty of Life Sciences and Advanced Materials and Nanotechnology Institute, Bar-Ilan University, Ramat-Gan 5290002, Israel.
[Ti] Título:The Canonical Poly (A) Polymerase PAP1 Polyadenylates Non-Coding RNAs and Is Essential for snoRNA Biogenesis in Trypanosoma brucei.
[So] Source:J Mol Biol;429(21):3301-3318, 2017 Oct 27.
[Is] ISSN:1089-8638
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The parasite Trypanosoma brucei is the causative agent of African sleeping sickness and is known for its unique RNA processing mechanisms that are common to all the kinetoplastidea including Leishmania and Trypanosoma cruzi. Trypanosomes possess two canonical RNA poly (A) polymerases (PAPs) termed PAP1 and PAP2. PAP1 is encoded by one of the only two genes harboring cis-spliced introns in this organism, and its function is currently unknown. In trypanosomes, all mRNAs, and non-coding RNAs such as small nucleolar RNAs (snoRNAs) and long non-coding RNAs (lncRNAs), undergo trans-splicing and polyadenylation. Here, we show that the function of PAP1, which is located in the nucleus, is to polyadenylate non-coding RNAs, which undergo trans-splicing and polyadenylation. Major substrates of PAP1 are the snoRNAs and lncRNAs. Under the silencing of either PAP1 or PAP2, the level of snoRNAs is reduced. The dual polyadenylation of snoRNA intermediates is carried out by both PAP2 and PAP1 and requires the factors essential for the polyadenylation of mRNAs. The dual polyadenylation of the precursor snoRNAs by PAPs may function to recruit the machinery essential for snoRNA processing.
[Mh] Termos MeSH primário: Poli A/genética
Poliadenilação/genética
Polinucleotídeo Adenililtransferase/genética
RNA Mensageiro/genética
RNA Nucleolar Pequeno/biossíntese
RNA não Traduzido/genética
Trypanosoma brucei brucei/enzimologia
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Proteínas Associadas a Pancreatite
Processamento de RNA
Alinhamento de Sequência
Trypanosoma brucei brucei/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Pancreatitis-Associated Proteins); 0 (REG3A protein, human); 0 (RNA, Messenger); 0 (RNA, Small Nucleolar); 0 (RNA, Untranslated); 24937-83-5 (Poly A); EC 2.7.7.19 (Polynucleotide Adenylyltransferase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:180116
[Lr] Data última revisão:
180116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE


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[PMID]:28977519
[Au] Autor:Han W; Pan S; López-Méndez B; Montoya G; She Q
[Ad] Endereço:Archaea Center, Department of Biology, University of Copenhagen, Ole Maal?es Vej 5, Copenhagen Biocenter, DK-2200 Copenhagen N, Denmark.
[Ti] Título:Allosteric regulation of Csx1, a type IIIB-associated CARF domain ribonuclease by RNAs carrying a tetraadenylate tail.
[So] Source:Nucleic Acids Res;45(18):10740-10750, 2017 Oct 13.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:CRISPR-Cas systems protect prokaryotes against invading viruses and plasmids. The system is associated with a large number of Cas accessory proteins among which many contain a CARF (CRISPR-associated Rossmann fold) domain implicated in ligand binding and a HEPN (higher eukaryotes and prokaryotes nucleotide-binding) nuclease domain. Here, such a dual domain protein, i.e. the Sulfolobus islandicus Csx1 (SisCsx1) was characterized. The enzyme exhibited metal-independent single-strand specific ribonuclease activity. In fact, SisCsx1 showed a basal RNase activity in the absence of ligand; upon the binding of an RNA ligand carrying four continuous adenosines at the 3'-end (3'-tetra-rA), the activated SisCsx1 degraded RNA substrate with a much higher turnover rate. Amino acid substitution mutants of SisCsx1 were obtained, and characterization of these mutant proteins showed that the CARF domain of the enzyme is responsible for binding to 3'-tetra-rA and the ligand binding strongly activates RNA cleavage by the HEPN domain. Since RNA polyadenylation is an important step in RNA decay in prokaryotes, and poly(A) RNAs can activate CARF domain proteins, the poly(A) RNA may function as an important signal in the cellular responses to viral infection and environmental stimuli, leading to degradation of both viral and host transcripts and eventually to cell dormancy or cell death.
[Mh] Termos MeSH primário: Proteínas Arqueais/metabolismo
Endorribonucleases/metabolismo
RNA Mensageiro/química
Sulfolobus/enzimologia
[Mh] Termos MeSH secundário: Regulação Alostérica
Proteínas Arqueais/química
Endorribonucleases/química
Ligantes
Metais/metabolismo
Poli A/química
Ligação Proteica
Domínios Proteicos
Clivagem do RNA
Sulfolobus/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Archaeal Proteins); 0 (Ligands); 0 (Metals); 0 (RNA, Messenger); 24937-83-5 (Poly A); EC 3.1.- (Endoribonucleases)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171005
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx726


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[PMID]:28911119
[Au] Autor:Huang C; Shi J; Guo Y; Huang W; Huang S; Ming S; Wu X; Zhang R; Ding J; Zhao W; Jia J; Huang X; Xiang AP; Shi Y; Yao C
[Ad] Endereço:Center for Stem Cell Biology and Tissue Engineering, Key Laboratory for Stem Cells and Tissue Engineering, Ministry of Education, Sun Yat-Sen University, Guangzhou 510080, China.
[Ti] Título:A snoRNA modulates mRNA 3' end processing and regulates the expression of a subset of mRNAs.
[So] Source:Nucleic Acids Res;45(15):8647-8660, 2017 Sep 06.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:mRNA 3' end processing is an essential step in gene expression. It is well established that canonical eukaryotic pre-mRNA 3' processing is carried out within a macromolecular machinery consisting of dozens of trans-acting proteins. However, it is unknown whether RNAs play any role in this process. Unexpectedly, we found that a subset of small nucleolar RNAs (snoRNAs) are associated with the mammalian mRNA 3' processing complex. These snoRNAs primarily interact with Fip1, a component of cleavage and polyadenylation specificity factor (CPSF). We have functionally characterized one of these snoRNAs and our results demonstrated that the U/A-rich SNORD50A inhibits mRNA 3' processing by blocking the Fip1-poly(A) site (PAS) interaction. Consistently, SNORD50A depletion altered the Fip1-RNA interaction landscape and changed the alternative polyadenylation (APA) profiles and/or transcript levels of a subset of genes. Taken together, our data revealed a novel function for snoRNAs and provided the first evidence that non-coding RNAs may play an important role in regulating mRNA 3' processing.
[Mh] Termos MeSH primário: Processamento de Terminações 3´ de RNA/genética
RNA Mensageiro/metabolismo
RNA Nucleolar Pequeno/fisiologia
[Mh] Termos MeSH secundário: Fator de Especificidade de Clivagem e Poliadenilação/metabolismo
Regulação da Expressão Gênica
Células HeLa
Seres Humanos
Proteínas Monoméricas de Ligação ao GTP/metabolismo
Poli A/metabolismo
Ligação Proteica
RNA Nucleolar Pequeno/metabolismo
Fatores de Poliadenilação e Clivagem de mRNA/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cleavage And Polyadenylation Specificity Factor); 0 (RNA, Messenger); 0 (RNA, Small Nucleolar); 0 (mRNA Cleavage and Polyadenylation Factors); 24937-83-5 (Poly A); EC 3.6.1.- (RRAGA protein, human); EC 3.6.5.2 (Monomeric GTP-Binding Proteins)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170916
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx651


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[PMID]:28610557
[Au] Autor:Bush SJ; McCulloch MEB; Summers KM; Hume DA; Clark EL
[Ad] Endereço:The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UK. stephen.bush@roslin.ed.ac.uk.
[Ti] Título:Integration of quantitated expression estimates from polyA-selected and rRNA-depleted RNA-seq libraries.
[So] Source:BMC Bioinformatics;18(1):301, 2017 Jun 13.
[Is] ISSN:1471-2105
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: The availability of fast alignment-free algorithms has greatly reduced the computational burden of RNA-seq processing, especially for relatively poorly assembled genomes. Using these approaches, previous RNA-seq datasets could potentially be processed and integrated with newly sequenced libraries. Confounding factors in such integration include sequencing depth and methods of RNA extraction and selection. Different selection methods (typically, either polyA-selection or rRNA-depletion) omit different RNAs, resulting in different fractions of the transcriptome being sequenced. In particular, rRNA-depleted libraries sample a broader fraction of the transcriptome than polyA-selected libraries. This study aimed to develop a systematic means of accounting for library type that allows data from these two methods to be compared. RESULTS: The method was developed by comparing two RNA-seq datasets from ovine macrophages, identical except for RNA selection method. Gene-level expression estimates were obtained using a two-part process centred on the high-speed transcript quantification tool Kallisto. Firstly, a set of reference transcripts was defined that constitute a standardised RNA space, with expression from both datasets quantified against it. Secondly, a simple ratio-based correction was applied to the rRNA-depleted estimates. The outcome is an almost perfect correlation between gene expression estimates, independent of library type and across the full range of levels of expression. CONCLUSION: A combination of reference transcriptome filtering and a ratio-based correction can create equivalent expression profiles from both polyA-selected and rRNA-depleted libraries. This approach will allow meta-analysis and integration of existing RNA-seq data into transcriptional atlas projects.
[Mh] Termos MeSH primário: Poli A/genética
RNA Ribossômico/genética
RNA/metabolismo
Análise de Sequência de RNA
Transcriptoma
[Mh] Termos MeSH secundário: Animais
Feminino
Perfilação da Expressão Gênica
Biblioteca Gênica
Lipopolissacarídeos/toxicidade
Macrófagos/citologia
Macrófagos/efeitos dos fármacos
Macrófagos/metabolismo
Masculino
RNA/química
RNA/isolamento & purificação
RNA Ribossômico/metabolismo
Ovinos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Lipopolysaccharides); 0 (RNA, Ribosomal); 24937-83-5 (Poly A); 63231-63-0 (RNA)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171102
[Lr] Data última revisão:
171102
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170615
[St] Status:MEDLINE
[do] DOI:10.1186/s12859-017-1714-9


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[PMID]:28559491
[Au] Autor:Chen CA; Zhang Y; Xiang Y; Han L; Shyu AB
[Ad] Endereço:Department of Biochemistry and Molecular Biology, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas 77030, USA.
[Ti] Título:Antagonistic actions of two human Pan3 isoforms on global mRNA turnover.
[So] Source:RNA;23(9):1404-1418, 2017 Sep.
[Is] ISSN:1469-9001
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Deadenylation is a fundamental process that regulates eukaryotic gene expression. Mammalian deadenylation exhibits biphasic kinetics, with the Pan2-Pan3 and Ccr4-Caf1 deadenylase complexes mediating the first and second phase, respectively; however, the significance of the biphasic nature of deadenylation in mRNA turnover remains unclear. In this study, we discovered that two distinct isoforms of human Pan3 display opposing properties necessary for coordinating the two phases of deadenylation. The shorter isoform (Pan3S) interacts more strongly with PABP than the longer isoform (Pan3L) does. Pan2 deadenylase activity is enhanced by Pan3S but suppressed by Pan3L. Knocking down individual Pan3 isoforms has opposing effects on the global poly(A) tail length profile, P-body formation, and different mRNA decay pathways. Transcriptome-wide analysis of Pan3 knockdown effects on mRNA turnover shows that depleting either Pan3 isoform causes profound and extensive changes in mRNA stability globally. These results reveal a new fundamental step governing mammalian mRNA metabolism. We propose that the first phase of deadenylation, coordinated through the interplay among the two Pan3 isoforms, Pan2, and PABP, represents a cytoplasmic mRNA maturation step important for proper mRNA turnover.
[Mh] Termos MeSH primário: Proteínas de Transporte/metabolismo
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
[Mh] Termos MeSH secundário: Animais
Proteínas de Transporte/química
Proteínas de Transporte/genética
Linhagem Celular
Proliferação Celular
Exorribonucleases/química
Exorribonucleases/metabolismo
Regulação da Expressão Gênica
Técnicas de Silenciamento de Genes
Seres Humanos
Camundongos
MicroRNAs/genética
Mutação
Poli A
Poliadenilação
Ligação Proteica
Domínios e Motivos de Interação entre Proteínas
Isoformas de Proteínas
Estabilidade de RNA
Transcriptoma
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Carrier Proteins); 0 (MicroRNAs); 0 (PAN3 protein, human); 0 (Protein Isoforms); 0 (RNA, Messenger); 24937-83-5 (Poly A); EC 3.1.- (Exoribonucleases); EC 3.1.13.4 (PAN2 protein, human)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171009
[Lr] Data última revisão:
171009
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170601
[St] Status:MEDLINE
[do] DOI:10.1261/rna.061556.117


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[PMID]:28537233
[Au] Autor:Ustyantsev IG; Golubchikova JS; Borodulina OR; Kramerov DA
[Ad] Endereço:Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, 119991 Russia.
[Ti] Título:[Canonical and noncanonical RNA polyadenylation].
[So] Source:Mol Biol (Mosk);51(2):262-273, 2017 Mar-Apr.
[Is] ISSN:0026-8984
[Cp] País de publicação:Russia (Federation)
[La] Idioma:rus
[Ab] Resumo:Polyadenylation is the non-template addition of adenosine nucleotides at the 3'-end of RNA, which occurs after transcription and generates a poly(A) tail up to 250-300 nucleotides long. In the first section of our review, we consider the classical process of mRNA 3'-terminus formation, which involves the cleavage of the transcript synthesized by RNA polymerase II and the associated poly(A) tail synthesis by canonical polyadenylate polymerase. Nucleotide sequences needed for mRNA cleavage and poly(A) tail synthesis, in particular the AAUAAA polyadenylation signal, as well as numerous proteins and their complexes involved in mRNA cleavage and polyadenylation, is described in detail. The significance of the poly(A) tail for prolonging mRNA lifetime and stimulating their translation is discussed. Data presented in the second section demonstrate that RNA transcribed by RNA polymerase III from certain SINEs (Short Interspersed Elements) can undergo AAUAAA-dependent polyadenylation. The structural and functional features of RNA polymerase III determine the unusual character of polyadenylation of RNAs synthesized by this enzyme. The history of recent developments in this area of study have been described in greater detail, in particular the discovery of AAUAAA-dependent polyadenylation of RNA synthesized by RNA polymerase III, which has not been discussed previously. Data on AAUAAA-independent polyadenylation catalyzed by noncanonical TRAMP poly(A)-polymerases (Trf4 and Trf5) have been presented in the third section. These enzymes promote rapid degradation of RNAs by adding a short poly(A) tail to them. This mechanism enables the recognition, poly(A)-marking, and elimination of incorrectly folded noncoding transcripts (e.g. ribosomal and transfer RNAs).
[Mh] Termos MeSH primário: Poli A/biossíntese
Poliadenilação/fisiologia
RNA Mensageiro/biossíntese
[Mh] Termos MeSH secundário: Animais
Seres Humanos
Poli A/genética
RNA Mensageiro/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (RNA, Messenger); 24937-83-5 (Poly A)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170814
[Lr] Data última revisão:
170814
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170525
[St] Status:MEDLINE
[do] DOI:10.7868/S0026898417010189


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[PMID]:28514447
[Au] Autor:Xu G; Greene GH; Yoo H; Liu L; Marqués J; Motley J; Dong X
[Ad] Endereço:Howard Hughes Medical Institute-Gordon and Betty Moore Foundation, Department of Biology, Duke University, Durham, North Carolina 27708, USA.
[Ti] Título:Global translational reprogramming is a fundamental layer of immune regulation in plants.
[So] Source:Nature;545(7655):487-490, 2017 05 25.
[Is] ISSN:1476-4687
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:In the absence of specialized immune cells, the need for plants to reprogram transcription to transition from growth-related activities to defence is well understood. However, little is known about translational changes that occur during immune induction. Using ribosome footprinting, here we perform global translatome profiling on Arabidopsis exposed to the microbe-associated molecular pattern elf18. We find that during this pattern-triggered immunity, translation is tightly regulated and poorly correlated with transcription. Identification of genes with altered translational efficiency leads to the discovery of novel regulators of this immune response. Further investigation of these genes shows that messenger RNA sequence features are major determinants of the observed translational efficiency changes. In the 5' leader sequences of transcripts with increased translational efficiency, we find a highly enriched messenger RNA consensus sequence, R-motif, consisting of mostly purines. We show that R-motif regulates translation in response to pattern-triggered immunity induction through interaction with poly(A)-binding proteins. Therefore, this study provides not only strong evidence, but also a molecular mechanism, for global translational reprogramming during pattern-triggered immunity in plants.
[Mh] Termos MeSH primário: Arabidopsis/genética
Arabidopsis/imunologia
Regulação da Expressão Gênica de Plantas
Padrões Moleculares Associados a Patógenos/imunologia
Imunidade Vegetal/genética
Biossíntese de Proteínas/genética
[Mh] Termos MeSH secundário: Sequência Consenso/genética
Perfilação da Expressão Gênica
Motivos de Nucleotídeos
Poli A/metabolismo
RNA Mensageiro/genética
RNA de Plantas/genética
Ribossomos/metabolismo
Transcrição Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Pathogen-Associated Molecular Pattern Molecules); 0 (RNA, Messenger); 0 (RNA, Plant); 24937-83-5 (Poly A)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:171117
[Lr] Data última revisão:
171117
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170518
[St] Status:MEDLINE
[do] DOI:10.1038/nature22371


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[PMID]:28490506
[Au] Autor:Nousch M; Minasaki R; Eckmann CR
[Ad] Endereço:Developmental Genetics, Institute of Biology, Martin Luther University Halle-Wittenberg, 06120 Halle (Saale), Germany.
[Ti] Título:Polyadenylation is the key aspect of GLD-2 function in .
[So] Source:RNA;23(8):1180-1187, 2017 Aug.
[Is] ISSN:1469-9001
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The role of many enzymes extends beyond their dedicated catalytic activity by fulfilling important cellular functions in a catalysis-independent fashion. In this aspect, little is known about 3'-end RNA-modifying enzymes that belong to the class of nucleotidyl transferases. Among these are noncanonical poly(A) polymerases, a group of evolutionarily conserved enzymes that are critical for gene expression regulation, by adding adenosines to the 3'-end of RNA targets. In this study, we investigate whether the functions of the cytoplasmic poly(A) polymerase (cytoPAP) GLD-2 in germ cells exclusively depend on its catalytic activity. To this end, we analyzed a specific missense mutation affecting a conserved amino acid in the catalytic region of GLD-2 cytoPAP. Although this mutated protein is expressed to wild-type levels and incorporated into cytoPAP complexes, we found that it cannot elongate mRNA poly(A) tails efficiently or promote GLD-2 target mRNA abundance. Furthermore, germ cell defects in animals expressing this mutant protein strongly resemble those lacking the GLD-2 protein altogether, arguing that only the polyadenylation activity of GLD-2 is essential for gametogenesis. In summary, we propose that all known molecular and biological functions of GLD-2 depend on its enzymatic activity, demonstrating that polyadenylation is the key mechanism of GLD-2 functionality. Our findings highlight the enzymatic importance of noncanonical poly(A) polymerases and emphasize the pivotal role of poly(A) tail-centered cytoplasmic mRNA regulation in germ cell biology.
[Mh] Termos MeSH primário: Proteínas de Caenorhabditis elegans/metabolismo
Caenorhabditis elegans/metabolismo
Regulação da Expressão Gênica
Poli A/metabolismo
Polinucleotídeo Adenililtransferase/metabolismo
Processamento Pós-Transcricional do RNA
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
Animais Geneticamente Modificados
Caenorhabditis elegans/genética
Caenorhabditis elegans/crescimento & desenvolvimento
Proteínas de Caenorhabditis elegans/genética
Mutação de Sentido Incorreto/genética
Poliadenilação
Polinucleotídeo Adenililtransferase/genética
Estabilidade de RNA
RNA Mensageiro/metabolismo
Alinhamento de Sequência
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Caenorhabditis elegans Proteins); 0 (RNA, Messenger); 24937-83-5 (Poly A); EC 2.7.7.19 (GLD-2 protein, C elegans); EC 2.7.7.19 (Polynucleotide Adenylyltransferase)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170922
[Lr] Data última revisão:
170922
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170512
[St] Status:MEDLINE
[do] DOI:10.1261/rna.061473.117


  10 / 8594 MEDLINE  
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[PMID]:28444238
[Au] Autor:Panda AC; De S; Grammatikakis I; Munk R; Yang X; Piao Y; Dudekula DB; Abdelmohsen K; Gorospe M
[Ad] Endereço:Laboratory of Genetics and Genomics, National Institute on Aging-Intramural Research Program, National Institutes of Health, Baltimore, MD 21224, USA.
[Ti] Título:High-purity circular RNA isolation method (RPAD) reveals vast collection of intronic circRNAs.
[So] Source:Nucleic Acids Res;45(12):e116, 2017 Jul 07.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:High-throughput RNA sequencing methods coupled with specialized bioinformatic analyses have recently uncovered tens of thousands of unique circular (circ)RNAs, but their complete sequences, genes of origin and functions are largely unknown. Given that circRNAs lack free ends and are thus relatively stable, their association with microRNAs (miRNAs) and RNA-binding proteins (RBPs) can influence gene expression programs. While exoribonuclease treatment is widely used to degrade linear RNAs and enrich circRNAs in RNA samples, it does not efficiently eliminate all linear RNAs. Here, we describe a novel method for the isolation of highly pure circRNA populations involving RNase R treatment followed by Polyadenylation and poly(A)+ RNA Depletion (RPAD), which removes linear RNA to near completion. High-throughput sequencing of RNA prepared using RPAD from human cervical carcinoma HeLa cells and mouse C2C12 myoblasts led to two surprising discoveries: (i) many exonic circRNA (EcircRNA) isoforms share an identical backsplice sequence but have different body sizes and sequences, and (ii) thousands of novel intronic circular RNAs (IcircRNAs) are expressed in cells. In sum, isolating high-purity circRNAs using the RPAD method can enable quantitative and qualitative analyses of circRNA types and sequence composition, paving the way for the elucidation of circRNA functions.
[Mh] Termos MeSH primário: Éxons
Sequenciamento de Nucleotídeos em Larga Escala
Íntrons
Poli A/genética
RNA Mensageiro/química
RNA/isolamento & purificação
[Mh] Termos MeSH secundário: Animais
Sequência de Bases
Linhagem Celular
Biologia Computacional
Exorribonucleases/química
Células HeLa
Seres Humanos
Camundongos
Anotação de Sequência Molecular
Mioblastos/citologia
Mioblastos/metabolismo
Poli A/metabolismo
Poliadenilação
RNA/genética
RNA/metabolismo
Clivagem do RNA
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Messenger); 0 (RNA, circular); 24937-83-5 (Poly A); 63231-63-0 (RNA); EC 3.1.- (Exoribonucleases); EC 3.1.27.- (ribonuclease R)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171011
[Lr] Data última revisão:
171011
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170427
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx297



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