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A Prunell

Publications and source records attributed to A Prunell.

At least 37 records · Page 2Linked to original sources
PubMed1986

Properties of supercoiled DNA in gel electrophoresis. The V-like dependence of mobility on topological constraint. DNA-matrix interactions.

The dependence of the electrophoretic mobility of small DNA rings on topological constraint was investigated in acrylamide or agarose gels as a function of DNA size (from approximately 350 to 1400 base-pairs), gel concentration and nucleotide sequence. Under appropriate adjustment between the size of the DNA and the gel concentration, this dependence was found to be V-shaped in a limited interval around constraint O, the minimum mobility at the apex of the V being obtained for relaxed DNA. Analysis of the DNA size dependence of the V suggests that it is the result of a modulated compaction of the DNA rings by the gel matrix. Compaction appears to be maximum upon relaxation, and to decrease with increase in supercoiling. Consistent with this interpretation, gels were found to oppose structural departures from the B helix, such as Z transition and cruciform extrusion, which tend to relax the DNA molecule and make it more expanded. In contrast, when DNA size or gel concentration are large enough relative to one another, U shapes are observed instead of Vs, as a consequence of an increase in the mobility of the rings closer to relaxation. The relevance of these results to the situation of superhelical DNA in vivo is discussed. Application of the V to the measurement of the DNA helical twist is mentioned.

Chemical Phenomena↗
PubMed1983

Periodicity of exonuclease III digestion of chromatin and the pitch of deoxyribonucleic acid on the nucleosome.

Exonuclease III has previously been shown to pause about every 10 nucleotides along the 3' strands while it invades the nucleosome core. Here, the exact periodicity of this digestion, i.e., the spacing of the pauses, was determined. Results showed that the exonuclease digests the first 20 nucleotides at the edge of the nucleosome core with a periodicity of approximately 11 nucleotides; in contrast, DNA closer to the center of the particle is digested with a smaller periodicity of about 10 nucleotides. These figures differ from the known periodicity of DNase I digestion, approximately 10 and 10.5 nucleotides at the edge and in the center of the nucleosome, respectively. Moreover, as shown by sedimentations in sucrose gradients, the structure of the nucleosome does not appear to be significantly altered by the gradual destruction of its DNA moiety by the exonuclease. Such stability of the nucleosome, along with other complementary observations, indicates that the transition in the digestion periodicity of the exonuclease may not be the consequence of a structural rearrangement of the particle upon trimming. This transition may rather be ascribed to the properties of the native nucleosome and to the intrinsic mechanism of action of the enzyme. Finally, evidence is presented which suggests that the exonuclease 10-nucleotide periodicity of digestion of the inner region of the nucleosome reflects a 10 base pair/turn pitch of the DNA in that region.

Animals↗
PubMed1983

Organization of internucleosomal DNA in rat liver chromatin.

A detailed analysis of the length distribution of DNA in nucleosome dimers trimmed with exonuclease III and S1 nuclease suggests that the previously described variation of internucleosomal distance in rat liver occurs, at least for a subset of the nucleosomes, by integral multiples of the helical repeat of the DNA. Results obtained upon digestion of chromatin with DNase II further suggest that lengths of internucleosomal DNA are integral multiples of the helical repeat of the DNA plus approximately 5 bp. Restraints imposed by these features on the arrangement of nucleosomes along the fiber are discussed.

Animals↗
PubMed1982

Nucleosome spacing in rat liver chromatin. A study with exonuclease III.

Exonuclease III was used to uniformly trim DNA ends of micrococcal nuclease-prepared chromatin fragments down to the first major impediment encountered by the enzyme, which arises from the interaction of H1 with the nucleosome. This trimming, when performed on nucleosome dimers, allowed one to quantitatively determine the center-to-center distance of nucleosomes. This distance, of mean 198 base pairs, was found to essentially vary between about 180 and 215 base pairs, with extremes of 165 and 230 base pairs. Trimming of trimers further revealed that the overall arrangement of nucleosome center-to-center distances along the chromatin fiber is that expected on a statistical basis.

Animals↗
PubMed1982

Nucleosome reconstitution on plasmid-inserted poly(dA) . poly(dT).

Chromatin was reconstituted from core histones and recombinant plasmid DNAs carrying poly(dA) . poly(dT) inserts of various lengths. A 97-bp insert was found to occupy discrete and regularly-spaced positions on the edges of the nucleosome. This insert cannot, however, be entirely included due to a block in the center of the particle. In contrast, nucleosomes reconstitute on a shorter 20-bp insert. In this case, the insert shows a marked preference for the edges of the particle. Possible structural and physiological implications of these observations are discussed.

Animals↗
PubMed1981

Helical periodicity of DNA, Poly(dA) . poly(dT) and poly(dA-dT). poly(dA-dT) in solution.

Helical periodicity of DNA, poly(dA) . poly(dT) and poly(dA-dT) . poly(dA-dT) has been measured in solution by using the band shift method fo Wang [Wang, J. (1979) Proc. Natl Acad. Sci. USA, 76, 200-203]. The method makes use of the effect, on the superhelicity of closed circular DNA molecules, of the insertion of specific nucleotide sequences of known length. The method was applied to a variety of recombinant plasmid DNAs which were constructed by inserting DNA, poly(dA) . poly(dT) or poly(dA-dT) . poly(dA-dT) into pBR322 DNA. When compared to DNA, poly(dA) . poly (dT) was found to have a smaller pitch (by about 0.5 base pair/turn), whereas poly(dA-dT) . poly(dA-dT) has a slightly larger pitch (by 0.1 base pair/turn). These features correlate well with the known ability of the alternating copolymer to reconstitute nucleosomes upon incubation with histones, in contrast to the non-alternating one which fails to do so. Finally, a detailed analysis of the principles underlying the methods developed by Wang [reference quoted above and Wang, J. (1978) Cold Spring Harb. Symp. Quant. Biol. 42, 29-33] leads to an increase in the estimate of the helical periodicity of DNA of 0.15 base pair/turn, over the reported value of 10.4 base pairs/turn (references quoted above). This essentially accounts for the discrepancy observed with the value of 10.6 base pairs/turn obtained by nuclease digestion of DNA immobilized on a surface [Rhodes, D & Klug, A. (1980) Nature (Lond.) 286, 573-578].

Base Sequence↗
PubMed1979

Periodicity of deoxyribonuclease I digestion of chromatin.

Two methods have been used to measure the single-strand lengths of the DNA fragments produced by deoxyribonuclease I digestion of chromatin. The average lengths obtained are muliples of about 10.4 bases, significantly different from the value of 10 previously reported. This periodicity in fragment lengths is closely related to the periodicity of the DNA double helix in chromatin, but the two values need not be exactly the same.

Animals↗
PubMed1978

Relation of nucleosomes to nucleotide sequences in the rat.

The relation of nucleosomes to nucleotide sequences is random for most single copy sequences in rat liver. This could be due to variation in the DNA content of nucleosomes, and a procedure for detecting such variation is described.

Animals↗