Substrate masking: binding of RNA by EGTA-inactivated micrococcal nuclease results in artifactual inhibition of RNA processing reactions.

Abstract:

:Inhibition of an RNA processing reaction after treatment with the Ca2(+)-dependent micrococcal nuclease (MN) is often used as a criterion for the presence of a required RNA or ribonucleoprotein component in the system. Following MN digestion, the nuclease is inactivated with EGTA and radiolabeled substrate is added to assay for remaining RNA processing activity. We found previously that inhibition of RNA processing by MN need not involve RNA hydrolysis: EGTA-inactivated MN can suppress RNA processing if the assay is performed in the absence of carrier RNA. We now demonstrate both by native gel electrophoresis and by nitrocellulose filter retention that EGTA-inactivated MN forms a complex with free RNA which can be dissociated by addition of synthetic polynucleotides or heparin. In the absence of Ca2+, nuclease binds to precursor tRNA with an apparent KD congruent to 1.4 x 10(-6) M, comparable to its reported affinity for DNA. In an assay for endonucleolytic tRNA maturation, inactivated MN bound to radiolabeled pre-tRNA physically blocks the sites of endonuclease cleavage and prevents tRNA processing. We call this phenomenon 'substrate masking'. Addition of excess carrier RNA competes with pre-tRNA for MN binding and restores normal processing.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Wang MJ,Gegenheimer P

doi

10.1093/nar/18.22.6625

subject

Has Abstract

pub_date

1990-11-25 00:00:00

pages

6625-31

issue

22

eissn

0305-1048

issn

1362-4962

journal_volume

18

pub_type

杂志文章
  • New tools and methods for direct programmatic access to the dbSNP relational database.

    abstract::Genome-wide association studies often incorporate information from public biological databases in order to provide a biological reference for interpreting the results. The dbSNP database is an extensive source of information on single nucleotide polymorphisms (SNPs) for many different organisms, including humans. We h...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq1054

    authors: Saccone SF,Quan J,Mehta G,Bolze R,Thomas P,Deelman E,Tischfield JA,Rice JP

    更新日期:2011-01-01 00:00:00

  • The transfer RNA of certain Enterobacteriacae contain 2-methylthiozeatin riboside (ms2io6A) an isopentenyl adenosine derivative.

    abstract::Isopentenyl adenosine derivatives are always located adjacent to the 3' end of the anticodon in transfer RNA and have been implicated in certain biological functions. In the enteric bacterium, E. coli, the derivative is ms2i6A whereas in some plant associated bacteria the derivative is the hydroxylated form, ms2io6A. ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/10.18.5663

    authors: Janzer JJ,Raney JP,McLennan BD

    更新日期:1982-09-25 00:00:00

  • Role of RNA structure in non-homologous recombination between genomic molecules of brome mosaic virus.

    abstract::Brome mosaic virus (BMV) is a tripartite genome, positive-sense RNA virus of plants. Previously it was demonstrated that local hybridization between BMV RNAs (RNA-RNA heteroduplex formation) efficiently promotes non-homologous RNA recombination. In addition, studies on the role of the BMV polymerase in RNA recombinati...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/28.8.1714

    authors: Figlerowicz M

    更新日期:2000-04-15 00:00:00

  • Transcription initiation sites within an IS2 insertion in a Gal-constitutive mutant of Escherichia coli.

    abstract::Insertion of the insertion sequence Is2(I) directly before the galE gene of the galactose operon results in a Gal minus phenotype (1, 2). The Gal-constitutive allele galc200 (and its deletion derivative galc200 delta 31) arise from such a Gal minus mutant by the insertion of LS2(II) DNA within the LS2(I) sequence (3)....

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/10.16.5015

    authors: Hinton DM,Musso RE

    更新日期:1982-08-25 00:00:00

  • NMR spectroscopy of the ring nitrogen protons of uracil and substituted uracils; relevance to A psi base pairing in the solution structure of transfer RNA.

    abstract::The chemical shifts of the well-resolved ring nitrogen protons of uracil and a series of substituted uracils, including pseudouridine (5-ribosyl-uracil) and uridine (1-ribosyl-uracil) were determined using nuclear magnetic resonance (NMR). The observed chemical shifts suggest the existence of an atypical syn conformat...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/4.8.2747

    authors: Hurd RE,Reid BR

    更新日期:1977-08-01 00:00:00

  • Chromosome XII context is important for rDNA function in yeast.

    abstract::The rDNA cluster in Saccharomyces cerevisiae is located 450 kb from the left end and 610 kb from the right end of chromosome XII and consists of approximately 150 tandemly repeated copies of a 9.1 kb rDNA unit. To explore the biological significance of this specific chromosomal context, chromosome XII was split at bot...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl293

    authors: Kim YH,Ishikawa D,Ha HP,Sugiyama M,Kaneko Y,Harashima S

    更新日期:2006-05-31 00:00:00

  • RNA aptamers selected against DNA polymerase beta inhibit the polymerase activities of DNA polymerases beta and kappa.

    abstract::DNA polymerase beta (polbeta), a member of the X family of DNA polymerases, is the major polymerase in the base excision repair pathway. Using in vitro selection, we obtained RNA aptamers for polbeta from a variable pool of 8 x 10(12) individual RNA sequences containing 30 random nucleotides. A total of 60 individual ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl326

    authors: Gening LV,Klincheva SA,Reshetnjak A,Grollman AP,Miller H

    更新日期:2006-05-17 00:00:00

  • Nematode.net update 2008: improvements enabling more efficient data mining and comparative nematode genomics.

    abstract::Nematode.net (http://nematode.net) is a publicly available resource dedicated to the study of parasitic nematodes. In 2000, the Genome Center at Washington University (GC) joined a consortium including the Nematode Genomics group in Edinburgh, and the Pathogen Sequencing Unit of the Sanger Institute to generate expres...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkn744

    authors: Martin J,Abubucker S,Wylie T,Yin Y,Wang Z,Mitreva M

    更新日期:2009-01-01 00:00:00

  • Alternative 5' exons either provide or deny an initiator methionine codon to the same alpha-tubulin coding region.

    abstract::The primary structures of two overlapping novel alpha-tubulin cDNA clones isolated from a Macaca fascicularis testis cDNA library and the corresponding human gene are presented. Although the general structure of the human gene conforms to that of previously described mammalian alpha-tubulin genes, there is a surprisin...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/15.1.199

    authors: Dobner PR,Kislauskis E,Wentworth BM,Villa-Komaroff L

    更新日期:1987-01-12 00:00:00

  • BTKbase, mutation database for X-linked agammaglobulinemia (XLA)

    abstract::X-linked agammaglobulinemia (XLA) is an immunodeficiency caused by mutations in the gene coding for Bruton's agammaglobulinemia tyrosine kinase (BTK). A database (BTKbase) of BTK mutations has been compiled and the recent update lists 368 entries from 318 unrelated families showing 228 unique molecular events. In addi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章,多中心研究

    doi:10.1093/nar/25.1.166

    authors: Vihinen M,Belohradsky BH,Haire RN,Holinski-Feder E,Kwan SP,Lappalainen I,Lehväslaiho H,Lester T,Meindl A,Ochs HD,Ollila J,Vorechovsky I,Weiss M,Smith CI

    更新日期:1997-01-01 00:00:00

  • Effects of Friedreich's ataxia (GAA)n*(TTC)n repeats on RNA synthesis and stability.

    abstract::Expansions of (GAA)n repeats within the first intron of the frataxin gene reduce its expression, resulting in a hereditary neurodegenerative disorder, Friedreich's ataxia. While it is generally believed that expanded (GAA)n repeats block transcription elongation, fine mechanisms responsible for gene repression are not...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl1140

    authors: Krasilnikova MM,Kireeva ML,Petrovic V,Knijnikova N,Kashlev M,Mirkin SM

    更新日期:2007-01-01 00:00:00

  • liqDB: a small-RNAseq knowledge discovery database for liquid biopsy studies.

    abstract::MiRNAs are important regulators of gene expression and are frequently deregulated under pathologic conditions. They are highly stable in bodily fluids which makes them feasible candidates to become minimally invasive biomarkers. In fact, several studies already proposed circulating miRNA-based biomarkers for different...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gky981

    authors: Aparicio-Puerta E,Jáspez D,Lebrón R,Koppers-Lalic D,Marchal JA,Hackenberg M

    更新日期:2019-01-08 00:00:00

  • Differential transcription of the orphan receptor RORbeta in nuclear extracts derived from Neuro2A and HeLa cells.

    abstract::An important model system for studying the process leading to productive transcription is provided by the superfamily of nuclear receptors, which are for the most part ligand-controlled transcription factors. Over the past years several 'orphan' nuclear receptors have been isolated for which no ligand has yet been ide...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/29.16.3424

    authors: Gawlas K,Stunnenberg HG

    更新日期:2001-08-15 00:00:00

  • Sequence complexity and diversity of polyadenylated RNA molecules of human normal resting, PHA-stimulated and leukemic lymphocytes.

    abstract::The abundance and complexity of total poly(A)+RNA of human normal unstimulated lymphocytes, PHA-stimulated and leukemic lymphocytes were examined using the cDNA-poly(A)+ RNA hybridization technique. It was found that in both PHA-stimulated and leukemic lymphocytes the complexity of total poly(A)+RNA is strongly reduce...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/9.24.7013

    authors: Torelli G,Ferrari S,Donelli A,Cadossi R,Ferrari S,Bosi P,Torelli U

    更新日期:1981-12-21 00:00:00

  • Thiostrepton-resistant mutants of Thermus thermophilus.

    abstract::Ribosomal protein L11 and its associated binding site on 23S rRNA together comprise one of the principle components that mediate interactions of translation factors with the ribosome. This site is also the target of the antibiotic thiostrepton, which has been proposed to act by preventing important structural transiti...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkh644

    authors: Cameron DM,Thompson J,Gregory ST,March PE,Dahlberg AE

    更新日期:2004-06-15 00:00:00

  • Checkpoint kinase 1 negatively regulates somatic hypermutation.

    abstract::Immunoglobulin (Ig) diversification by somatic hypermutation in germinal center B cells is instrumental for maturation of the humoral immune response, but also bears the risk of excessive or aberrant genetic changes. Thus, introduction of DNA damage by activation-induced cytidine deaminase as well as DNA repair by mul...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkt1378

    authors: Frankenberger S,Davari K,Fischer-Burkart S,Böttcher K,Tomi NS,Zimber-Strobl U,Jungnickel B

    更新日期:2014-04-01 00:00:00

  • Molecular structure of a U•A-U-rich RNA triple helix with 11 consecutive base triples.

    abstract::Three-dimensional structures have been solved for several naturally occurring RNA triple helices, although all are limited to six or fewer consecutive base triples, hindering accurate estimation of global and local structural parameters. We present an X-ray crystal structure of a right-handed, U•A-U-rich RNA triple he...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkz1222

    authors: Ruszkowska A,Ruszkowski M,Hulewicz JP,Dauter Z,Brown JA

    更新日期:2020-04-06 00:00:00

  • Structural repeat units of Chinese hamster ovary chromatin. Evidence for variations in repeat unit DNA size in higher eukaryotes.

    abstract::DNA lengths in the structural repeat units of Chinese hamster ovary (CHO) and chicken erythrocyte chromatin were compared by analyzing the sizes of DNA fragments produced after treatment of nuclei with staphylococcal nuclease. The repeat length of CHO chromatin (173 +- 4 BP) is about 20 base pairs (BP) smaller than th...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/4.4.771

    authors: Rill RL,Nelson DA,Oosterhof DK,Hozier JC

    更新日期:1977-04-01 00:00:00

  • Complementation of aprataxin deficiency by base excision repair enzymes in mitochondrial extracts.

    abstract::Mitochondrial aprataxin (APTX) protects the mitochondrial genome from the consequence of ligase failure by removing the abortive ligation product, i.e. the 5'-adenylate (5'-AMP) group, during DNA replication and repair. In the absence of APTX activity, blocked base excision repair (BER) intermediates containing the 5'...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx654

    authors: Çaglayan M,Prasad R,Krasich R,Longley MJ,Kadoda K,Tsuda M,Sasanuma H,Takeda S,Tano K,Copeland WC,Wilson SH

    更新日期:2017-09-29 00:00:00

  • PathPred: an enzyme-catalyzed metabolic pathway prediction server.

    abstract::The KEGG RPAIR database is a collection of biochemical structure transformation patterns, called RDM patterns, and chemical structure alignments of substrate-product pairs (reactant pairs) in all known enzyme-catalyzed reactions taken from the Enzyme Nomenclature and the KEGG PATHWAY database. Here, we present PathPre...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq318

    authors: Moriya Y,Shigemizu D,Hattori M,Tokimatsu T,Kotera M,Goto S,Kanehisa M

    更新日期:2010-07-01 00:00:00

  • DNA damage-induced activation of CUL4B targets HUWE1 for proteasomal degradation.

    abstract::The E3 ubiquitin ligase HUWE1/Mule/ARF-BP1 plays an important role in integrating/coordinating diverse cellular processes such as DNA damage repair and apoptosis. A previous study has shown that HUWE1 is required for the early step of DNA damage-induced apoptosis, by targeting MCL-1 for proteasomal degradation. Howeve...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkv325

    authors: Yi J,Lu G,Li L,Wang X,Cao L,Lin M,Zhang S,Shao G

    更新日期:2015-05-19 00:00:00

  • SpliceInfo: an information repository for mRNA alternative splicing in human genome.

    abstract::We have developed an information repository named SpliceInfo to collect the occurrences of the four major alternative-splicing (AS) modes in human genome; these include exon skipping, 5'-alternative splicing, 3'-alternative splicing and intron retention. The dataset is derived by comparing the nucleotide and protein s...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gki129

    authors: Huang HD,Horng JT,Lin FM,Chang YC,Huang CC

    更新日期:2005-01-01 00:00:00

  • The elongation factor 1 A-2 isoform from rabbit: cloning of the cDNA and characterization of the protein.

    abstract::Eukaryotic elongation factor 1 A (eEF1A, formerly elongation factor-1 alpha) is an important component of the protein synthesis apparatus. Here we report the isolation and characterization of the cDNA sequence encoding rabbit eEF1A-2, an isoform of eEF1A, as well as a structural and functional comparison of the two ra...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/26.8.1884

    authors: Kahns S,Lund A,Kristensen P,Knudsen CR,Clark BF,Cavallius J,Merrick WC

    更新日期:1998-04-15 00:00:00

  • On the importance of the primer activation signal for initiation of tRNA(lys3)-primed reverse transcription of the HIV-1 RNA genome.

    abstract::Initiation of reverse transcription is a complex and regulated process in all retroviruses. Several base pairing interactions have been proposed to occur between the HIV-1 RNA genome and the specific tRNA(lys3) primer. The tRNA primer can form up to 21 bp with the primer binding site (PBS), and an additional 8 bp inte...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkg714

    authors: Huthoff H,Bugala K,Barciszewski J,Berkhout B

    更新日期:2003-09-01 00:00:00

  • The signal recognition particle database (SRPDB).

    abstract::The SRPDB (signal recognition particle database) provides aligned SRP RNA and protein sequences, annotated and phylogenetically ordered. This release includes 82 SRP RNAs (including 22 bacterial and 9 archaeal homologs) and a total of 20 protein sequences representing SRP9, SRP14, SRP19, SRP54, SRP68, and SRP72. The o...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/22.17.3483

    authors: Zwieb C,Larsen N

    更新日期:1994-09-01 00:00:00

  • New efficient sulfurizing reagents for the preparation of oligodeoxyribonucleotide phosphorothioate analogues.

    abstract::A set of new sulfurizing agents representing disulfides of arylsulfonic acids has been developed for the automated synthesis of phosphorothioate oligonucleotide analogues via the phosphoramidite method. These reagents, such as bis(benzenesulfonyl)disulfide, bis(p-toluenesulfonyl)disulfide, bis(p-methoxybenzensulfonyl)...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.20.4029

    authors: Efimov VA,Kalinkina AL,Chakhmakhcheva OG,Hill TS,Jayaraman K

    更新日期:1995-10-25 00:00:00

  • Premature termination of RNA polymerase II mediated transcription of a seed protein gene in Schizosaccharomyces pombe.

    abstract::The poly(A) signal and downstream elements with transcriptional pausing activity play an important role in termination of RNA polymerase II transcription. We show that an intronic sequence derived from the plant seed protein gene (AmA1) specifically acts as a transcriptional terminator in the fission yeast, Schizosacc...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkf334

    authors: Chakraborty S,Sarmah B,Chakraborty N,Datta A

    更新日期:2002-07-01 00:00:00

  • Cap-independent translation initiation in Xenopus oocytes.

    abstract::Eukaryotic cellular mRNAs contain a cap at their 5'-ends, but some viral and cellular mRNAs bypass the cap-dependent mechanism of translation initiation in favor of internal entry of ribosomes at specific RNA sequences. Cap-dependent initiation requires intact initiation factor eIF4G (formerly eIF-4gamma, eIF-4Fgamma ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/25.2.395

    authors: Keiper BD,Rhoads RE

    更新日期:1997-01-15 00:00:00

  • Cloning and characterization of the MboII restriction-modification system.

    abstract::The two genes encoding the class IIS restriction-modification system MboII from Moraxella bovis were cloned separately in two compatible plasmids and expressed in E. coli RR1 delta M15. The nucleotide sequences of the MboII endonuclease (R.MboII) and methylase (M.MboII) genes were determined and the putative start cod...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/19.5.1007

    authors: Bocklage H,Heeger K,Müller-Hill B

    更新日期:1991-03-11 00:00:00

  • SynergyFinder 2.0: visual analytics of multi-drug combination synergies.

    abstract::SynergyFinder (https://synergyfinder.fimm.fi) is a stand-alone web-application for interactive analysis and visualization of drug combination screening data. Since its first release in 2017, SynergyFinder has become a widely used web-tool both for the discovery of novel synergistic drug combinations in pre-clinical mo...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkaa216

    authors: Ianevski A,Giri AK,Aittokallio T

    更新日期:2020-07-02 00:00:00