The structure of histone-depleted metaphase chromosomes

Paulson, James R.; Laemmli, Ulrich Karl

doi:10.1016/0092-8674(77)90280-x

Cited by 1,062 publications

(584 citation statements)

References 18 publications

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“…Electron microscopic and sedimentation analyses of histone-depleted chromosomes and nuclei (5,14,40,52) support the hypothesis that eucaryotic DNA is organized as individual loops constrained at their bases by interaction of DNA with nonhistone proteins. Laemmli and co-workers (37) have proposed an extraction procedure in which lithium 3'-5'-diiodosalicylate (LIS) is used to isolate specific DNA fragments which presumably form the bases of the loops.…”

mentioning

confidence: 53%

Studies of an 800-kilobase DNA stretch of the Drosophila X chromosome: comapping of a subclass of scaffold-attached regions with sequences able to replicate autonomously in Saccharomyces cerevisiae.

Brun¹,

Dang²,

Miassod³

1990

Mol. Cell. Biol.

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We have previously mapped scaffold-attached regions (SARs) on an 800-kilobase DNA walk from the Drosophila X chromosome. We have also previously shown that the strength of binding, i.e., the ability of SARs to bind to all nuclear scaffolds or only to a fraction of them varied from one SAR to another one. In the present study, 71 of the 85 subfragments that bind scaffolds and 38 fragments that do not bind scaffolds were tested for their ability to promote autonomous replicating sequence (ARS) activity in Saccharomyces cerevisiae. Sixteen SAR-containing fragments from the chromosome walk were also examined for association to yeast nuclear scaffolds in vitro. All identified ARSs (a total of 27) were present on SAR-containing fragments, except two, which were adjacent to SARs. There is thus a correlation between ARS and SAR activities, and this correlation defines a SAR subclass. Moreover, the presence of an ARS on a DNA fragment appeared to be highly correlated with the strength of binding. The binding activity was highly conserved from Drosophila melanogaster to yeast. These data suggest that Drosophila DNA sequences responsible for binding to components of the nuclear scaffold from either D. melanogaster or yeast may be involved in the process of heterologous extrachromosomal replication in yeasts.Electron microscopic and sedimentation analyses of histone-depleted chromosomes and nuclei (5, 14, 40, 52) support the hypothesis that eucaryotic DNA is organized as individual loops constrained at their bases by interaction of DNA with nonhistone proteins. Laemmli and co-workers (37) have proposed an extraction procedure in which lithium 3'-5'-diiodosalicylate (LIS) is used to isolate specific DNA fragments which presumably form the bases of the loops. This technique has been applied to various parts of the

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mentioning

confidence: 53%

Studies of an 800-kilobase DNA stretch of the Drosophila X chromosome: comapping of a subclass of scaffold-attached regions with sequences able to replicate autonomously in Saccharomyces cerevisiae.

Brun¹,

Dang²,

Miassod³

1990

Mol. Cell. Biol.

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“…One of these structures is the chromosome scaffold, a structural framework that organizes along mitotic chromosomes (Paulson and Laemmli 1977) and whose main components are DNA topoisomerase IIα and the condensin subunit SMC2 (Earnshaw et al 1985;Saitoh et al 1994). Because DNA topoisomerase IIα is associated to the LEs during meiosis, it has been proposed that the scaffold and the SC may have some degree of interaction (Moens and Earnshaw 1989).…”

Section: Introductionmentioning

confidence: 99%

Sex chromosomes, synapsis, and cohesins: a complex affair

et al. 2006

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During first meiotic prophase, homologous chromosomes are held together by the synaptonemal complex, a tripartite proteinaceous structure that extends along the entire length of meiotic bivalents. While this feature is applicable for autosomes, sex chromosomes often escape from this rule. Many species present sex chromosomes that differ between them in their morphology, length, and gene content. Moreover, in some species, sex chromosomes appear in a single dose in one of the sexes. In all of these cases, the behavior of sex chromosomes during meiosis is conspicuously affected, and this includes the assembly and dynamics of the synaptonemal complex. We review in this study the structure of the synaptonemal complex in the sex chromosomes of three groups of organisms, namely: mammals, orthopterans, and hemipterans, which present different patterns of sex chromosome structure and behavior. Of special interest is the analysis of the organization of the axial/lateral elements of the synaptonemal complex in relation to other axial structures organized along meiotic chromosomes, mainly the cohesin axis. The differences found in the behavior of both axial structures reveal that while the organization of a cohesin axis along sex chromosomes is a conserved feature in most organisms and it shows very little morphological variations, the axial/lateral elements of the synaptonemal complex present a wide range of structural modifications on these chromosomes.

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“…Our studies with mitotic chromosomes are particularly revealing because it was possible to study the histone-depleted chromosomes both biochemically and microscopically (3,4).…”

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confidence: 99%

Isolation of a protein scaffold from mitotic HeLa cell chromosomes

Adolph

Cheng

Paulson

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

169

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We have recently shown that, after the histones and most of the nonhistone proteins are gently removed from HeLa metaphase chromosomes, the chromosomal DNA is still highly organized and relatively compact. The structure of these histone-depleted chromosomes is due to the presence of a number of nonhistone proteins that form a central scaffold that retains the approximate size and shape of intact chromosomes and to which the DNA is attached, predominantly forming loops. We now demonstrate that the protein scaffold may be iso ated independently of the DNA by treating HeLa chromosomes with micrococcal nuclease before removing the histones.The chromosomal scaffolds may be isolated by sucrose density gradient centrifugation as a well-defined peak that is stable in 2 M sodium chloride, but is dissociated by treatment with proteases, 4 M urea, or 0.1% sodium dodecyl sulfate. Polyacrylamide gel electrophoresis reveals that the protein content of scaffold preparations is identical to that of histone-depleted chromosomes. Fluorescence microscopy of purified scaffolds in isolation buffer shows that the particles still possess the familiar chromosome morphology. When the scaffolds are examined in the electron microscope, a fibrous structure with the approximate size and shape of intact, paired chromatids is seen. Less than 0.1I% of the chromosomal DNA and virtually no histones are associated with the purified scaffold structures. Mitotic chromosomes are composed of the five histones and a large number of nonhistone proteins that keep the DNA in a highly compact arrangement. Electron microscopy has revealed that the nucleoprotein fiber in mitotic chromosomes is a "knobby" fiber with a diameter of about 200-300 A and appears in a complex, net-like arrangement (e.g., ref. 1). The complexity of this network and the tendency of the chromatin fibers to adhere to each other make it difficult to study the higher-order folding of the nucleoprotein fiber in chromosomes.A great deal has been learned in the last few years about the structural role played by the histones in chromatin, but little is known about a possible involvement of nonhistone proteins in chromosome structure. Starting with the hypothesis that nonhistone proteins-may play a role in determining the superstructure of interphase and metaphase chromosomes, we have recently developed an approach that permits dissection of the structural contribution made by the histones and certain nonhistone proteins. Our general approach toward uncovering a structural role for the nonhistone proteins has been to gently remove the histones from chromosomes by competition with dextran sulfate and heparin, without destroying the continuity of the DNA fiber (2). In this way, all the histones and many nonhistone proteins are removed, leaving behind some tightly bound nonhistone proteins that keep the chromosomal DNA highly organized and compact. This dissecting procedure hasThe costs of publication of this article were defrayed in part by the payment of page charges. This article mu...

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The structure of histone-depleted metaphase chromosomes

Cited by 1,062 publications

References 18 publications

Studies of an 800-kilobase DNA stretch of the Drosophila X chromosome: comapping of a subclass of scaffold-attached regions with sequences able to replicate autonomously in Saccharomyces cerevisiae.

Studies of an 800-kilobase DNA stretch of the Drosophila X chromosome: comapping of a subclass of scaffold-attached regions with sequences able to replicate autonomously in Saccharomyces cerevisiae.

Sex chromosomes, synapsis, and cohesins: a complex affair

Isolation of a protein scaffold from mitotic HeLa cell chromosomes

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