Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.30.1 (S1 nuclease)
 3,660 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

S1 nuclease hypersensitive sites in the 5' flanking regions of eukaryotic genes, present in small artificial supercoiled DNA circles, reside in homopurine-homopyrimidine stretches. The hierarchical behavior which these sites exhibit is consistent with the notion that they act as sinks of torsional free energy. By employing DMS as a single-strand-specific reagent, we show that these sites (despite their S1 sensitivity) are regions of duplex DNA. A simple thermodynamic treatment indicates that the high torsional stress in the small DNA circles is almost certain to be relieved by the formation of alternate DNA structures. The same treatment places some constraints on the types and sizes of the regions with alternate conformation. While no definitive structural conclusions can be drawn, left-handed helices seem most consistent with the extent and the pattern of sensitivity to S1 nuclease.
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PMID:Possible structures of homopurine-homopyrimidine S1-hypersensitive sites. 609 87

A synthetic deoxyoligonucleotide containing five palindromic repeats of GGATCC self assembles to form a parallel four-stranded structure held together by G-tetrads that shows slower mobility than duplex DNA. This structure is hypersensitive to S1 nuclease and resistant to DMS modification. The same oligonucleotide when cloned in a plasmid forms a different structure under supercoiling that persists stably even in the cleaved out insert. On polyacrylamide gel electrophoresis, the cleaved out insert moves to a position midway between the duplex and parallel four-stranded forms of the oligonucleotide. Upon S1 nuclease treatment, the cleaved out insert shows a discreet band of 18 base pairs, suggesting an unfolded region in the middle. All the guanines in the cleaved out insert are sensitive to DMS modification and produce a positive peak at 285 nM in the circular dichroism spectrum, a signature of fold back tetraplex structures. We propose a fold back quadruplex structure for the insert under supercoiling with only A.T.A.T and G.C.G.C tetrads. This is the first suggestive evidence of a general tetraplex motif without G quartets as that proposed for generalized recombination.
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PMID:A palindromic repeat sequence adopts a stable fold back structure under supercoiling. 1642 17

Abstract Formation of triple helices with GA and GT third strands has been studied. Besides the usual investigation techniques common for characterizing triple helical formation (CD spectroscopy, gel shift mobility assay, chemical probing and S1 nuclease footprinting) we have used a new technique in which targeting of polypurine sequences in duplexes was demonstrated on oligonucleotide microchips. This technique is very successful to quickly test a large number of potential triple helix formation. In this work we used oligonucleotide microairay to study the specificity of DNA duplex recognition by GA and GT strands. Generic 6-mer microchip containing all possible 4(6) = 4,096 single-stranded hexadeoxyribonucleotides immobilized within individual gel pads was applied. To study formation of intermolecular triple helices on the generic microchip, a number of Pu.Py duplexes were formed by hybridization of the mixture of purine octadeoxyribonucleotides on the microchip followed by targeting of the duplexes by GA or GT third strands. Melting behavior of the formed structures was investigated using fluorescence measurements under microscope. In solution we present the results obtained for GT triplexes and discuss the characteristics of the CD spectra. Results obtained by S1 nuclease footprinting, KMnO(4) and DMS chemical probing are consistent with the spectroscopic data reflecting triplex structure formation.
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PMID:Targeting of Pu.Py Duplexes by GA and GT Rich Oligonucleotides on Microchip and in Solution. 2260 29

Guanine-rich DNAs can fold into four-stranded structures that contain stacks of G-quartets. Bioinformatics studies have revealed that G-rich sequences with the potential to adopt these structures are unevenly distributed throughout genomes, and are especially found in gene promoter regions. With the exception of the single-stranded telomeric DNA, all genomic G-rich sequences will always be present along with their C-rich complements, and quadruplex formation will be in competition with the corresponding Watson-Crick duplex. Quadruplex formation must therefore first require local dissociation (melting) of the duplex strands. Since negative supercoiling is known to facilitate the formation of alternative DNA structures, we have investigated G-quadruplex formation within negatively supercoiled DNA plasmids. Plasmids containing multiple copies of (G3T)n and (G3T4)n repeats, were probed with dimethylsulphate, potassium permanganate and S1 nuclease. While dimethylsulphate footprinting revealed some evidence for G-quadruplex formation in (G3T)n sequences, this was not affected by supercoiling, and permanganate failed to detect exposed thymines in the loop regions. (G3T4)n sequences were not protected from DMS and showed no reaction with permanganate. Similarly, both S1 nuclease and 2D gel electrophoresis of DNA topoisomers did not detect any supercoil-dependent structural transitions. These results suggest that negative supercoiling alone is not sufficient to drive G-quadruplex formation.
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PMID:The effects of DNA supercoiling on G-quadruplex formation. 2903 19