Abstract:
:Bleomycin B(2)(BLM) in the presence of iron [Fe(II)] and O(2)catalyzes single-stranded (ss) and double-stranded (ds) cleavage of DNA. Electrospray ionization ion trap mass spectrometry was used to monitor these cleavage processes. Two duplex oligonucleotides containing an ethylene oxide tether between both strands were used in this investigation, allowing facile monitoring of all ss and ds cleavage events. A sequence for site-specific binding and cleavage by Fe-BLM was incorporated into each analyte. One of these core sequences, GTAC, is a known hot-spot for ds cleavage, while the other sequence, GGCC, is a hot-spot for ss cleavage. Incubation of each oligo-nucleotide under anaerobic conditions with Fe(II)-BLM allowed detection of the non-covalent ternary Fe-BLM/oligonucleotide complex in the gas phase. Cleavage studies were then performed utilizing O(2)-activated Fe(II)-BLM. No work-up or separation steps were required and direct MS and MS/MS analyses of the crude reaction mixtures confirmed sequence-specific Fe-BLM-induced cleavage. Comparison of the cleavage patterns for both oligonucleotides revealed sequence-dependent preferences for ss and ds cleavages in accordance with previously established gel electrophoresis analysis of hairpin oligonucleotides. This novel methodology allowed direct, rapid and accurate determination of cleavage profiles of model duplex oligonucleotides after exposure to activated Fe-BLM.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Harsch A,Marzilli LA,Bunt RC,Stubbe J,Vouros Pdoi
10.1093/nar/28.9.1978keywords:
subject
Has Abstractpub_date
2000-05-01 00:00:00pages
1978-85issue
9eissn
0305-1048issn
1362-4962pii
gkd308journal_volume
28pub_type
杂志文章abstract::Monocrotaline is a pyrrolizidine alkaloid known to cause toxicity in humans and animals. Its mechanism of biological action is still unclear although DNA crosslinking has been suggested to a play a role in its activity. In this study we found that an active metabolite of monocrotaline, dehydromonocrotaline (DHM), alky...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.23.5441
更新日期:1998-12-01 00:00:00
abstract::A stable conformer of Escherichia coli tRNA(Glu), obtained in the absence of Mg(2+), is inactive in the aminoacylation reaction. Probing it with diethylpyrocarbonate, dimethyl sulfate and ribonuclease V1 revealed that it has a hairpin structure with two internal loops; the helical segments at both extremities have the...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/27.17.3583
更新日期:1999-09-01 00:00:00
abstract::Approximately 2% of the Dictyostelium discoideum genome consists of multiple copies of a retrotransposable element termed DRE (Dictyostelium Repetitive Element). These elements have always been found integrated in a position and orientation-specific manner 50 +/- 4 nucleotides upstream of the coding region of tRNA gen...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/20.23.6247
更新日期:1992-12-11 00:00:00
abstract::Analysis of 41 histone homologous clones from an isogenic gene library of Drosophila melanogaster showed that non-histone fragments interrupt the histone repetitive clusters at several sites. Long (L) and short (S) forms of the repeating units are distinguished by the insertion of 240 bp into the spacer between H1 and...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/17.1.225
更新日期:1989-01-11 00:00:00
abstract::We have examined the interactions of UvrABC endonuclease with DNA containing the monoadducts of 8-methoxypsoralen (8-MOP) and 4,5',8-trimethylpsoralen (TMP). The UvrA and UvrB proteins were found to form a stable complex on DNA that contains the psoralen monoadducts. Subsequent binding of UvrC protein to this complex ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.12.4957
更新日期:1987-06-25 00:00:00
abstract::Molecular Biology Database List (MBDL) includes brief descriptions and pointers to Web sites for the various databases described in this issue as well as other Web sites presenting data sets relevant to molecular biology. This information is compiled into a list (http://www.oup.co.uk/nar/Volume_27/Issue_01/summary/ gk...
journal_title:Nucleic acids research
pub_type:
doi:10.1093/nar/27.1.1
更新日期:1999-01-01 00:00:00
abstract::Calf lens messenger fractions have been purified on poly(dT)-cellulose. The purified RNA's have been analyzed as nucleosides by a method which can be used on a micro-scale with highly reproduceble results. Both the 10S and 14S lens mRNA contain poly A sequences. The poly A content found in 14S mRNA cannot account for ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/1.2.245
更新日期:1974-02-01 00:00:00
abstract::Over millions of years the structure and complexity of the transcriptional regulatory network (TRN) in bacteria has changed, reorganized and enabled them to adapt to almost every environmental niche on earth. In order to understand the plasticity of TRNs in bacteria, we studied the conservation of currently known TRNs...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkl423
更新日期:2006-07-13 00:00:00
abstract::Sequencing of RNAs (RNA-Seq) has revolutionized the field of transcriptomics, but the reads obtained often contain errors. Read error correction can have a large impact on our ability to accurately assemble transcripts. This is especially true for de novo transcriptome analysis, where a reference genome is not availab...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkt215
更新日期:2013-05-01 00:00:00
abstract::In this report we have analyzed the role of antisense transcription in the control of LEF1 transcription factor expression. A natural antisense transcript (NAT) is transcribed from a promoter present in the first intron of LEF1 gene and undergoes splicing in mesenchymal cells. Although this locus is silent in epitheli...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv502
更新日期:2015-07-13 00:00:00
abstract::The structure of eukaryotic chromatin has been investigated by isolating and analyzing the "accessible" DNA fraction of rat liver chromatin. This DNA fraction has been isolated by titrating the chromatin with the protese-resistant D isomer of polylysine to bind the "accessible" DNA sites. After removal of chromosomal ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/2.5.667
更新日期:1975-05-01 00:00:00
abstract::Contrary to the expectation, the Merrifield polystyrene resin, 2% cross-linked by divinylbenzene, is as efficient as the polyacrylmorpholide resin for the synthesis of polydeoxyribonucleotides using a phosphotriester method. On the Merrifield resin, the tetradecamer, dTpCpGpTpCpApApCpTpGpGpCpTpT, and the hexadecamer, ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/8.22.5507
更新日期:1980-11-25 00:00:00
abstract::We analyzed the effects of site-directed mutations in the SUC2 promoter of Saccharomyces cerevisiae. Analyses were performed in wild-type as well as mig1 and tup1 mutant strains after the promoter mutants were reintroduced into the native SUC2 locus on the left arm of chromosome IX. Mutation of the two GC boxes reveal...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/26.4.1002
更新日期:1998-02-15 00:00:00
abstract::Work from a number of laboratories has indicated that the TATA box sequence can act as a basal promoter element not only for RNA polymerase II (RNAP II) transcription, but also for transcription by RNA polymerase III (RNAP III). We previously reported that, in the absence of other cis-acting elements, the canonical TA...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/24.15.3100
更新日期:1996-08-01 00:00:00
abstract::Restriction endonuclease Bse634I recognizes and cleaves the degenerate DNA sequence 5'-R/CCGGY-3' (R stands for A or G; Y for T or C, '/' indicates a cleavage position). Here, we report the crystal structures of the Bse634I R226A mutant complexed with cognate oligoduplexes containing ACCGGT and GCCGGC sites, respectiv...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gks300
更新日期:2012-08-01 00:00:00
abstract::The somatic V(D)J recombination for the assembly of the Ig and TCR genes is mediated by the recombination signal sequences (Rss) and the V(D)J recombinase. A cDNA clone was isolated from a lambda gt11 expression library made from mouse thymocyte poly(A)+ RNA, using the Rss as a ligand. The deduced amino acid sequence ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/21.22.5067
更新日期:1993-11-11 00:00:00
abstract::Genome organization into transcriptionally active domains denotes one of the first levels of gene expression regulation. Although the chromatin domain concept is generally accepted, only little is known on how domain organization impacts the regulation of differential gene expression. Insulators might hold answers to ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq740
更新日期:2011-01-01 00:00:00
abstract::The Agrobacterium tumefaciens Ti plasmid contains at least six transcriptional units (designated vir loci) which are essential for efficient crown gall tumorigenesis. Mutations in one of these loci, virE, result in a sharply attenuated virulence phenotype. In the present communication, we have analyzed the virE operon...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/15.2.825
更新日期:1987-01-26 00:00:00
abstract::Smarcal1 is a SWI/SNF-family protein with an ATPase domain involved in DNA-annealing activities and a binding site for the RPA single-strand-DNA-binding protein. Although the role played by Smarcal1 in the maintenance of replication forks has been established, it remains unknown whether Smarcal1 contributes to genomic...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkv621
更新日期:2015-07-27 00:00:00
abstract::CAG repeats occur predominantly in the coding regions of human genes, which suggests their functional importance. In some genes, these sequences can undergo pathogenic expansions leading to neurodegenerative polyglutamine (poly-Q) diseases. The mutant transcripts containing expanded CAG repeats possibly contribute to ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkq700
更新日期:2010-12-01 00:00:00
abstract::The human Microprocessor complex cleaves primary microRNA (miRNA) transcripts (pri-miRNAs) to initiate miRNA synthesis. Microprocessor consists of DROSHA (an RNase III enzyme), and DGCR8. DROSHA contains two RNase III domains, RIIIDa and RIIIDb, which simultaneously cleave the 3p- and 5p-strands of pri-miRNAs, respect...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa018
更新日期:2020-03-18 00:00:00
abstract::Based on the secondary structural model of MS2 RNA, it is shown that, in base-pairing regions of the RNA, there is a bias in the use of synonymous codons which favours C and/or G over U and/or A in the third codon positions, and that in non-pairing regions, there is an opposite bias which favours U and/or A over C and...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/7.7.2073
更新日期:1979-12-11 00:00:00
abstract::Maintaining genome integrity is important for cells and damaged DNA triggers autoimmunity. Previous studies have reported that Three-prime repair exonuclease 1(TREX1), an endogenous DNA exonuclease, prevents immune activation by depleting damaged DNA, thus preventing the development of certain autoimmune diseases. Con...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkx178
更新日期:2017-05-05 00:00:00
abstract::Prostatic steroid binding protein, whose expression is stimulated by androgens, consists of two subunits: one containing the polypeptides C1 and C3 and the other containing C2 and C3. We have characterised genomic clones containing the C1 and C2 genes by restriction enzyme analysis and DNA sequencing. Both genes are 3...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/10.17.5121
更新日期:1982-09-11 00:00:00
abstract::RNA structural complexity and flexibility present a challenge for computational modeling efforts. Experimental information and bioinformatics data can be used as restraints to improve the accuracy of RNA tertiary structure prediction. Regarding utilization of restraints, the fundamental questions are: (i) What is the ...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkz427
更新日期:2019-06-20 00:00:00
abstract::Prokaryotes evolved numerous systems that defend against predation by bacteriophages. In addition to well-known restriction-modification and CRISPR-Cas immunity systems, many poorly characterized systems exist. One class of such systems, named BREX, consists of a putative phosphatase, a methyltransferase and four othe...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gky1125
更新日期:2019-01-10 00:00:00
abstract::The determination of distances between specific points in nucleic acids is essential to understanding their behaviour at the molecular level. The ability to measure distances of 2-10 nm is particularly important: deformations arising from protein binding commonly fall within this range, but the reliable measurement of...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa086
更新日期:2020-04-06 00:00:00
abstract::Cdc48/p97 is an evolutionary conserved ubiquitin-dependent chaperone involved in a broad array of cellular functions due to its ability to associate with multiple cofactors. Aside from its role in removing RNA polymerase II from chromatin after DNA damage, little is known about how this AAA-ATPase is involved in the t...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gku786
更新日期:2014-01-01 00:00:00
abstract::Chromosome conformation-capture technologies are widely used in 3D genomics; however, experimentally, such methods have high-noise limitations and, therefore, require significant bioinformatics efforts to extract reliable distal interactions. Miscellaneous undesired linear DNAs, present during proximity-ligation, repr...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa106
更新日期:2020-05-07 00:00:00
abstract::Inducible promoters are a central regulatory component in synthetic biology, metabolic engineering, and protein production for laboratory and commercial uses. Many of these applications utilize two or more exogenous promoters, imposing a currently unquantifiable metabolic burden on the living system. Here, we engineer...
journal_title:Nucleic acids research
pub_type: 杂志文章
doi:10.1093/nar/gkaa734
更新日期:2020-09-25 00:00:00