An improved method for photofootprinting yeast genes in vivo using Taq polymerase.

Abstract:

:We have developed an improved method for photofootprinting in vivo which utilizes the thermostable DNA polymerase from T. aquaticus (Taq) in a primer extension assay. UV light is used to introduce photoproducts into the genomic DNA of intact yeast cells. The photoproducts are then detected and mapped at the nucleotide level by multiple rounds of annealing and extension using Taq polymerase, which is blocked by photoproducts in the template DNA. The method is more rapid, sensitive, and reproducible than the previously described chemical photofootprinting procedure developed in this laboratory (Nature 325. 173-177), and detects photoproducts with a specificity which is similar, but not identical to that of the previously described procedure. Binding of GAL4 protein to its binding sites within the GAL1-10 upstream activating sequence is demonstrated using the primer extension photofootprinting method. The primer extension assay can also be used to map DNA strand breakage generated by other footprinting methods, and to determine DNA sequence directly from the yeast genome.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Axelrod JD,Majors J

doi

10.1093/nar/17.1.171

subject

Has Abstract

pub_date

1989-01-11 00:00:00

pages

171-83

issue

1

eissn

0305-1048

issn

1362-4962

journal_volume

17

pub_type

杂志文章
  • The nucleotide sequence of the ubiquitous repetitive DNA sequence B1 complementary to the most abundant class of mouse fold-back RNA.

    abstract::Three copies of a highly repetitive DNA sequence B1 which is complementary to the most abundant class of mouse fold-back RNA have been cloned in pBR322 plasmid and sequenced by the method of Maxam and Gilbert. All the three have a length of about 130 base pairs and are very similar in their base sequence. The deviatio...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/8.6.1201

    authors: Krayev AS,Kramerov DA,Skryabin KG,Ryskov AP,Bayev AA,Georgiev GP

    更新日期:1980-03-25 00:00:00

  • WormNet v3: a network-assisted hypothesis-generating server for Caenorhabditis elegans.

    abstract::High-throughput experimental technologies gradually shift the paradigm of biological research from hypothesis-validation toward hypothesis-generation science. Translating diverse types of large-scale experimental data into testable hypotheses, however, remains a daunting task. We previously demonstrated that heterogen...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gku367

    authors: Cho A,Shin J,Hwang S,Kim C,Shim H,Kim H,Kim H,Lee I

    更新日期:2014-07-01 00:00:00

  • Structure determination of a nucleoside Q precursor isolated from E. coli tRNA: 7-(aminomethyl)-7-deazaguanosine.

    abstract::A precursor of modified nucleoside Q isolated from E. coli methyl-deficient tRNA was determined to be 7-(aminomethyl)-7-deazaguanosine. The structure was deduced by means of its chromatographic and electrophoretic mobilities, and UV and mass spectra, in addition to comparison with the synthesized authentic compound. T...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/5.7.2289

    authors: Okada N,Noguchi S,Nishimura S,Ohgi T,Goto T,Crain PF,McCloskey JA

    更新日期:1978-07-01 00:00:00

  • Interactions between subunits of Saccharomyces cerevisiae RNase MRP support a conserved eukaryotic RNase P/MRP architecture.

    abstract::Ribonuclease MRP is an endonuclease, related to RNase P, which functions in eukaryotic pre-rRNA processing. In Saccharomyces cerevisiae, RNase MRP comprises an RNA subunit and ten proteins. To improve our understanding of subunit roles and enzyme architecture, we have examined protein-protein and protein-RNA interacti...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkm553

    authors: Aspinall TV,Gordon JM,Bennett HJ,Karahalios P,Bukowski JP,Walker SC,Engelke DR,Avis JM

    更新日期:2007-01-01 00:00:00

  • The ovalbumin gene family: complete sequence and structure of the Y gene.

    abstract::The "ovalbumin Y" gene, one of three which constitute the ovalbumin gene family in chicken has been completely sequenced. The exact location of exons can be derived from the comparison with the ovalbumin gene sequence and from the map previously established by electron microscopy analysis. During evolution of the Y ge...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/10.14.4363

    authors: Heilig R,Muraskowsky R,Kloepfer C,Mandel JL

    更新日期:1982-07-24 00:00:00

  • Ligation of oligonucleotides to nucleic acids or proteins via disulfide bonds.

    abstract::We have developed general methods for joining together, via cleavable disulfide bonds, either two unprotected polynucleotides or a polynucleotide and a peptide or protein. To join two oligonucleotides, each is first converted to an adduct in which cystamine is joined to the 5'-terminal phosphate of the oligonucleotide...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/16.9.3671

    authors: Chu BC,Orgel LE

    更新日期:1988-05-11 00:00:00

  • The genomic organisation and nucleotide sequence of the HLA-SB(DP) alpha gene.

    abstract::We have isolated a unique fragment of the HLA-DR alpha gene and probed human genomic DNA at low stringency to search for homologous sequences. A minimum of six non-polymorphic cross-hybridizing high molecular weight fragments were found in all DNAs examined. In order to obtain molecular clones of these cross-hybridizi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/13.20.7515

    authors: Lawrance SK,Das HK,Pan J,Weissman SM

    更新日期:1985-10-25 00:00:00

  • Interdependence between DNA template secondary structure and priming efficiencies of short primers.

    abstract::Here we analyze the effect of DNA folding on the performance of short primers and describe a simple technique for assessing hitherto uncertain values of thermodynamic parameters that determine the folding of single-stranded DNA into secondary structure. An 8mer with two degenerate positions is extended simultaneously ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/26.23.5525

    authors: Lvovsky L,Ioshikhes I,Raja MC,Zevin-Sonkin D,Sobolev IA,Liberzon A,Shwartzburd J,Ulanovsky LE

    更新日期:1998-12-01 00:00:00

  • Combined transcriptome and proteome analysis as a powerful approach to study genes under glucose repression in Bacillus subtilis.

    abstract::We used 2D protein gel electrophoresis and DNA microarray technologies to systematically analyze genes under glucose repression in B:acillus subtilis. In particular, we focused on genes expressed after the shift from glycolytic to gluconeogenic at the middle logarithmic phase of growth in a nutrient sporulation medium...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/29.3.683

    authors: Yoshida K,Kobayashi K,Miwa Y,Kang CM,Matsunaga M,Yamaguchi H,Tojo S,Yamamoto M,Nishi R,Ogasawara N,Nakayama T,Fujita Y

    更新日期:2001-02-01 00:00:00

  • 5-methylcytosine in RNA: detection, enzymatic formation and biological functions.

    abstract::The nucleobase modification 5-methylcytosine (m(5)C) is widespread both in DNA and different cellular RNAs. The functions and enzymatic mechanisms of DNA m(5)C-methylation were extensively studied during the last decades. However, the location, the mechanism of formation and the cellular function(s) of the same modifi...

    journal_title:Nucleic acids research

    pub_type: 杂志文章,评审

    doi:10.1093/nar/gkp1117

    authors: Motorin Y,Lyko F,Helm M

    更新日期:2010-03-01 00:00:00

  • Monitoring the spatio-temporal organization and dynamics of the genome.

    abstract::The spatio-temporal organization of chromatin in the eukaryotic cell nucleus is of vital importance for transcription, DNA replication and genome maintenance. Each of these activities is tightly regulated in both time and space. While we have a good understanding of chromatin organization in space, for example in fixe...

    journal_title:Nucleic acids research

    pub_type: 杂志文章,评审

    doi:10.1093/nar/gkaa135

    authors: Shaban HA,Seeber A

    更新日期:2020-04-17 00:00:00

  • On the cooperative and noncooperative binding of ethidium to DNA.

    abstract::The equilibrium binding of ethidium bromide to native DNAs and to poly(dG-dC) X poly(dG-dC) has been studied by both phase partition and direct spectrophotometric techniques. The binding isotherms obtained from both experimental techniques show that ethidium binds in a cooperative manner to E. coli DNA. On the other h...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/10.24.8211

    authors: Winkle SA,Rosenberg LS,Krugh TR

    更新日期:1982-12-20 00:00:00

  • BRENDA in 2015: exciting developments in its 25th year of existence.

    abstract::The BRENDA enzyme information system (http://www.brenda-enzymes.org/) has developed into an elaborate system of enzyme and enzyme-ligand information obtained from different sources, combined with flexible query systems and evaluation tools. The information is obtained by manual extraction from primary literature, text...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gku1068

    authors: Chang A,Schomburg I,Placzek S,Jeske L,Ulbrich M,Xiao M,Sensen CW,Schomburg D

    更新日期:2015-01-01 00:00:00

  • The non-autonomous retrotransposon SVA is trans-mobilized by the human LINE-1 protein machinery.

    abstract::SINE-VNTR-Alu (SVA) elements are non-autonomous, hominid-specific non-LTR retrotransposons and distinguished by their organization as composite mobile elements. They represent the evolutionarily youngest, currently active family of human non-LTR retrotransposons, and sporadically generate disease-causing insertions. S...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr863

    authors: Raiz J,Damert A,Chira S,Held U,Klawitter S,Hamdorf M,Löwer J,Strätling WH,Löwer R,Schumann GG

    更新日期:2012-02-01 00:00:00

  • Enhancing multiplex genome editing by natural transformation (MuGENT) via inactivation of ssDNA exonucleases.

    abstract::Recently, we described a method for multiplex genome editing by natural transformation (MuGENT). Mutant constructs for MuGENT require large arms of homology (>2000 bp) surrounding each genome edit, which necessitates laborious in vitro DNA splicing. In Vibrio cholerae, we uncover that this requirement is due to cytopl...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkx496

    authors: Dalia TN,Yoon SH,Galli E,Barre FX,Waters CM,Dalia AB

    更新日期:2017-07-07 00:00:00

  • The MH1 domain of Smad3 interacts with Pax6 and represses autoregulation of the Pax6 P1 promoter.

    abstract::Pax6 transcription is under the control of two main promoters (P0 and P1), and these are autoregulated by Pax6. Additionally, Pax6 expression is under the control of the TGFbeta superfamily, although the precise mechanisms of such regulation are not understood. The effect of TGFbeta on Pax6 expression was studied in t...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl1105

    authors: Grocott T,Frost V,Maillard M,Johansen T,Wheeler GN,Dawes LJ,Wormstone IM,Chantry A

    更新日期:2007-01-01 00:00:00

  • Tight cooperation between Mot1p and NC2β in regulating genome-wide transcription, repression of transcription following heat shock induction and genetic interaction with SAGA.

    abstract::TATA-binding protein (TBP) is central to the regulation of eukaryotic transcription initiation. Recruitment of TBP to target genes can be positively regulated by one of two basal transcription factor complexes: SAGA or TFIID. Negative regulation of TBP promoter association can be performed by Mot1p or the NC2 complex....

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr784

    authors: Spedale G,Meddens CA,Koster MJ,Ko CW,van Hooff SR,Holstege FC,Timmers HT,Pijnappel WW

    更新日期:2012-02-01 00:00:00

  • Next-generation RNA-based fluorescent biosensors enable anaerobic detection of cyclic di-GMP.

    abstract::Bacteria occupy a diverse set of environmental niches with differing oxygen availability. Anaerobic environments such as mammalian digestive tracts and industrial reactors harbor an abundance of both obligate and facultative anaerobes, many of which play significant roles in human health and biomanufacturing. Studying...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw580

    authors: Wang XC,Wilson SC,Hammond MC

    更新日期:2016-09-30 00:00:00

  • Molecular footprints of human immunoglobulin gene evolution: a new sequence family.

    abstract::Analysis of the human VK (ref. 2) gene locus led to the detection of a new sequence family (L sequences). Its copy number is in the range of 10(2). The L sequences, which are about 500 bp long, are found as part of the 3' flanking regions of a clustered set of human VKI genes but they occur also separate from the gene...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/12.13.5265

    authors: Straubinger B,Pech M,Mühlebach K,Jaenichen HR,Bauer HG,Zachau HG

    更新日期:1984-07-11 00:00:00

  • DIANA-microT Web server upgrade supports Fly and Worm miRNA target prediction and bibliographic miRNA to disease association.

    abstract::microRNAs (miRNAs) are small endogenous RNA molecules that are implicated in many biological processes through post-transcriptional regulation of gene expression. The DIANA-microT Web server provides a user-friendly interface for comprehensive computational analysis of miRNA targets in human and mouse. The server has ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkr294

    authors: Maragkakis M,Vergoulis T,Alexiou P,Reczko M,Plomaritou K,Gousis M,Kourtis K,Koziris N,Dalamagas T,Hatzigeorgiou AG

    更新日期:2011-07-01 00:00:00

  • O-ribosyl-phosphate purine as a constant modified nucleotide located at position 64 in cytoplasmic initiator tRNAs(Met) of yeasts.

    abstract::The unknown modified nucleotide G*, isolated from both Schizosaccharomyces pombe and Torulopsis utilis initiator tRNAs(Met), has been identified as an O-ribosyl-(1"----2')-guanosine-5"-phosphate, called Gr(p), by means of HPLC, UV-absorption, mass spectrometry and periodate oxidation procedures. By comparison with the...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/19.19.5199

    authors: Glasser AL,Desgres J,Heitzler J,Gehrke CW,Keith G

    更新日期:1991-10-11 00:00:00

  • Site-specific modification of the lactose operator with acetylaminofluorene.

    abstract::We have synthesized the tetradecamer GAGCXGATAACAAG containing a part of the sequence of the lactose operator. A guanine base in the sequence is replaced by the adduct of the carcinogen 2-acetylaminofluorene with guanine. Under the standard conditions of de-protection, the fluorene moiety is lost, leaving behind a gua...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/11.15.5093

    authors: Stöhrer G,Osband JA,Alvarado-Urbina G

    更新日期:1983-08-11 00:00:00

  • 20-Hydroxyecdysone increases levels of transient gene expression in transfected Drosophila cells.

    abstract::Methods for increasing the transient level of expression of transfected DNA in cultured Drosophila cells have been examined. Here we show that cells exposed to the steroid hormone 20-hydroxyecdysone for 48h prior to transfection show an 4 to 5 fold increase in the levels of expression of transfected DNA. By analysis, ...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/15.22.9255

    authors: Sinclair JH,Bryant LA

    更新日期:1987-11-25 00:00:00

  • GeneAlign: a coding exon prediction tool based on phylogenetical comparisons.

    abstract::GeneAlign is a coding exon prediction tool for predicting protein coding genes by measuring the homologies between a sequence of a genome and related sequences, which have been annotated, of other genomes. Identifying protein coding genes is one of most important tasks in newly sequenced genomes. With increasing numbe...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkl307

    authors: Hsieh SJ,Lin CY,Liu NH,Chow WY,Tang CY

    更新日期:2006-07-01 00:00:00

  • Nucleosome-free DNA regions differentially affect distant communication in chromatin.

    abstract::Communication between distantly spaced genomic regions is one of the key features of gene regulation in eukaryotes. Chromatin per se can stimulate efficient enhancer-promoter communication (EPC); however, the role of chromatin structure and dynamics in this process remains poorly understood. Here we show that nucleoso...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkw1240

    authors: Nizovtseva EV,Clauvelin N,Todolli S,Polikanov YS,Kulaeva OI,Wengrzynek S,Olson WK,Studitsky VM

    更新日期:2017-04-07 00:00:00

  • The role of TFIIB-RNA polymerase II interaction in start site selection in yeast cells.

    abstract::Previous studies have established a critical role of both TFIIB and RNA polymerase II (RNAPII) in start site selection in the yeast Saccharomyces cerevisiae. However, it remains unclear how the TFIIB-RNAPII interaction impacts on this process since such an interaction can potentially influence both preinitiation compl...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkf422

    authors: Zhang DY,Carson DJ,Ma J

    更新日期:2002-07-15 00:00:00

  • Vitamin D receptor contains multiple dimerization interfaces that are functionally different.

    abstract::The vitamin D receptor mediates the signal of 1 alpha, 25-dihydroxyvitamin D3 by binding to vitamin D responsive elements in DNA as a homodimer or as a heterodimer composed of one vitamin D receptor subunit and one retinoid X receptor subunit. We have mapped the dimerization interfaces of the vitamin D receptor that i...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/23.4.606

    authors: Nishikawa J,Kitaura M,Imagawa M,Nishihara T

    更新日期:1995-02-25 00:00:00

  • Germline ablation of SMUG1 DNA glycosylase causes loss of 5-hydroxymethyluracil- and UNG-backup uracil-excision activities and increases cancer predisposition of Ung-/-Msh2-/- mice.

    abstract::Deamination of cytosine (C), 5-methylcytosine (mC) and 5-hydroxymethylcytosine (hmC) occurs spontaneously in mammalian DNA with several hundred deaminations occurring in each cell every day. The resulting potentially mutagenic mispairs of uracil (U), thymine (T) or 5-hydroxymethyluracil (hmU) with guanine (G) are subs...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks259

    authors: Kemmerich K,Dingler FA,Rada C,Neuberger MS

    更新日期:2012-07-01 00:00:00

  • EMdeCODE: a novel algorithm capable of reading words of epigenetic code to predict enhancers and retroviral integration sites and to identify H3R2me1 as a distinctive mark of coding versus non-coding genes.

    abstract::Existence of some extra-genetic (epigenetic) codes has been postulated since the discovery of the primary genetic code. Evident effects of histone post-translational modifications or DNA methylation over the efficiency and the regulation of DNA processes are supporting this postulation. EMdeCODE is an original algorit...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gks1214

    authors: Santoni FA

    更新日期:2013-02-01 00:00:00

  • T1DBase: update 2011, organization and presentation of large-scale data sets for type 1 diabetes research.

    abstract::T1DBase (http://www.t1dbase.org) is web platform, which supports the type 1 diabetes (T1D) community. It integrates genetic, genomic and expression data relevant to T1D research across mouse, rat and human and presents this to the user as a set of web pages and tools. This update describes the incorporation of new dat...

    journal_title:Nucleic acids research

    pub_type: 杂志文章

    doi:10.1093/nar/gkq912

    authors: Burren OS,Adlem EC,Achuthan P,Christensen M,Coulson RM,Todd JA

    更新日期:2011-01-01 00:00:00