Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold

Mirkovitch, Jovan; Mirault, Marc-Édouard; Laemmli, Ulrich Karl

doi:10.1016/0092-8674(84)90208-3

Cited by 1,067 publications

(824 citation statements)

References 45 publications

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“…Isolation of LIS-generated halo structures The procedure used is a modification of the protocol of Dijkwel and Hamlin (1988); the LIS technique was originally developed by Mirkovitch et al (1984). All procedures were carried out at 4°C unless indicated otherwise.…”

Section: Dna Transfectionmentioning

confidence: 99%

“…The approach taken here employs the lithium diiodosalicylate (LIS) procedure (Mirkovitch et al, 1984 (Mirkovitch et al, 1984;Cockerill and Garrard, 1986;Amati and Gasser, 1988;see Gasser et al, 1989 for review (Moyzis et al, 1988;Allshire et al, 1989;Cross et al, 1989;de Lange et al, 1990). In most DNA preparations TTAGGG repeat probes hybridize to heterogeneously sized restriction fragments with an average molecular weight that varies in different cell lines and tissues.…”

Section: Introductionmentioning

confidence: 99%

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Human telomeres are attached to the nuclear matrix.

1992

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This report shows that human telomeres are tightly associated with the nuclear matrix. Telomere attachment is observed in several cell types and in all stages of interphase. Mapping experiments show that telomeres are anchored via their TTAGGG repeats; a subtelomeric repeat located immediately proximal to the telomeric TTAGGG repeats is quantitatively released from the nuclear matrix by restriction endonuclease cleavage. TTAGGG repeats introduced at chromosome-internal sites by DNA transfection do not behave as matrix attached loci, suggesting that the telomeric position of the repeats is required for their interaction with the nuclear matrix. These findings are consistent with the idea that telomeres function as a nucleoprotein complex.

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Section: Dna Transfectionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Human telomeres are attached to the nuclear matrix.

1992

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“…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 Drosophila melanogaster (18,19,26,36,37; J. Mirkovitch, P. Spierer, and U. K. Laemmli, Letter, J. Mol. Biol.…”

mentioning

confidence: 99%

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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“…The feasibility of these preparations for FISH analysis was shown using adherently growing fibroblasts . The extraction procedure results in extended DNA loops that surround a matrix fraction that is thought to represent the nuclear scaffold (Vogelstein et al 1980;Mirkovitch et al 1984). We have prepared Halos from lymphoblastoid cell lines of a carrier of DMD.…”

Section: Discussionmentioning

confidence: 99%

Multicolor fluorescence in situ hybridization on metaphase chromosomes and interphase Halo-preparations using cosmid and YAC clones for the simultaneous high resolution mapping of deletions in the dystrophin gene

et al. 1994

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Abstract. We report on multicolor fluorescence in situ hybridization protocols for the simultaneous visualization of deletion-prone regions for carrier detection of Duchenne/ Becker (DMD/BMD) muscular dystrophy. Cosmid and yeast artificial chromosome (YAC) clones specific for preferentially deleted subregions of the dystrophin gene were labeled differentially and detected with three different fluorochromes using digital imaging microscopy. This approach allows for an assessment of the carrier status of female relatives even in families where no index patient is available. Cosmid and YAC clones, and different probegeneration protocols are compared with respect to their feasibility for cartier detection. The use of histone-depleted interphase nuclei (Halo-preparations) for deletion mapping is demonstrated and shown to have a resolution power of 5 kb.

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Organization of the higher-order chromatin loop: specific DNA attachment sites on nuclear scaffold

Cited by 1,067 publications

References 45 publications

Human telomeres are attached to the nuclear matrix.

Human telomeres are attached to the nuclear matrix.

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.

Multicolor fluorescence in situ hybridization on metaphase chromosomes and interphase Halo-preparations using cosmid and YAC clones for the simultaneous high resolution mapping of deletions in the dystrophin gene

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