Chromatin Structure in Bands and Interbands of Polytene Chromosomes Imaged by Atomic Force Microscopy

Grauw, C.J. de; Avogadro, A.; Heuvel, D.J. van den; Werf, Kees O. van der; Otto, Cees; Kraan, Yvonne M.; Hulst, Niek F. van; Greve, Jan

doi:10.1006/jsbi.1997.3929

Cited by 12 publications

(8 citation statements)

References 28 publications

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“…A closer examination of immunostained chromosomes revealed that Dnmts were always associated with interbands of polytene chromosomes ( Figures 2C-2E). Interbands represent polytene chromosomal areas that are less intensely stained by 4,6diamidino-2-phenylindole and contain less-condensed chromatin fibres [31]. In addition, interbands have also been shown to contain hypoacetylated histones and the SIN3-RPD3 histone deacetylase complex [32].…”

Section: Resultsmentioning

confidence: 99%

“…Analysis of immunostained polytene chromosomes revealed a consistent localization of Dnmts to interbands. While the function of polytene chromosome banding patterns remains to be resolved, interbands have been shown to contain relatively less condensed chromatin [31]. An interband-specific polytene chromosome staining pattern association has also been observed for hypoacetylated histones H3 and H4 and for the SIN3-RPD3 histone deacetylase complex and has been interpreted to represent a specific marker of repressed genes in transcriptionally active regions [32].…”

Section: Discussionmentioning

confidence: 98%

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Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases

Mund

Musch

Strödicke

et al. 2004

Biochemical Journal

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DNA methyltransferases (Dnmts) mediate the epigenetic modification of eukaryotic genomes. Mammalian DNA methylation patterns are established and maintained by co-operative interactions among the Dnmt proteins Dnmt1, Dnmt3a and Dnmt3b. Owing to their simultaneous presence in mammalian cells, the activities of individual Dnmt have not yet been determined. This includes a fourth putative Dnmt, namely Dnmt2, which has failed to reveal any activity in previous assays. We have now established transgenic Drosophila strains that allow for individual overexpression of all known mouse Dnmts. Quantitative analysis of genomic cytosine methylation levels demonstrated a robust Dnmt activity for the de novo methyltransferases Dnmt3a and Dnmt3b. In addition, we also detected a weak but significant activity for Dnmt2. Subsequent methylation tract analysis by genomic bisulphite sequencing revealed that Dnmt3 enzymes preferentially methylated CpG dinucleotides in a processive manner, whereas Dnmt2 methylated isolated cytosine residues in a non-CpG dinucleotide context. Our results allow a direct comparison of the activities of mammalian Dnmts and suggest a significant functional specialization of these enzymes.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 98%

Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases

Mund

Musch

Strödicke

et al. 2004

Biochemical Journal

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“…DAPI staining is weak or absent in interbands and puffs. Chromatin condensation levels inferred from DAPI staining intensity are supported by atomic force microscopy, which reveals that interbands contain chromatin fibers that range from 11 to 300 nm, while bands contain predominantly 700 nm fibers (de Grauw et al ., 1998).…”

Section: The Sin3–rpd3 Complex Binds Less‐condensed Interbands and Ismentioning

confidence: 99%

Chromosomal localization links the SIN3-RPD3 complex to the regulation of chromatin condensation, histone acetylation and gene expression

Pile

Wassarman

2000

The EMBO Journal

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Acetylation of core histone N-terminal tails influences chromatin condensation and transcription. To examine how the SIN3-RPD3 deacetylase complex contributes to these events in vivo, we examined binding of SIN3 and RPD3 to Drosophila salivary gland polytene chromosomes. The binding patterns of SIN3 and RPD3 were highly coincident, suggesting that the SIN3-RPD3 complex is the most abundant chromatin-bound RPD3 complex in salivary gland cells. SIN3- RPD3 binding was restricted to less condensed, hypoacetylated euchromatic interbands and was absent from moderately condensed, hyperacetylated euchromatic bands and highly condensed, differentially acetylated centric heterochromatin. Consistent with its demonstrated role in transcriptional repression, SIN3-RPD3 did not co-localize with RNA polymer ase II. Chromatin binding of the complex, mediated by SMRTER, decreased upon ecdysone-induced transcriptional activation but was restored when transcription was reduced. These results implicate SIN3-RPD3 in maintaining histone acetylation levels or patterns within less condensed chromatin domains and suggest that SIN3-RPD3 activity is required, in the absence of an activation signal, to repress transcription of particular genes within transcriptionally active chromatin domains.

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“…Ultrastructural studies using mostly electron microscopy (EM) and recently atomic force microscopy (AFM) reveal a series of fiber structures including 11-nm, 30-nm and at the higher level, 250-nm and 700-nm fibers in polytene chromosomes, indicating a general pattern of packaging similar to that found in mitotic chromosomes de Grauw et al, 1998). It was reported that F-actin is directly associated with polytene chromosomes and essential for their structural maintenance (Sauman and Berry, 1994).…”

Section: Literature Review Chromatin Structure and Organizationmentioning

confidence: 89%

Remodeling of higher order chromatin structure by the JIL-1 histone H3 kinase in Drosophila

Huai

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Chromatin Structure in Bands and Interbands of Polytene Chromosomes Imaged by Atomic Force Microscopy

Cited by 12 publications

References 28 publications

Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases

Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases

Chromosomal localization links the SIN3-RPD3 complex to the regulation of chromatin condensation, histone acetylation and gene expression

Remodeling of higher order chromatin structure by the JIL-1 histone H3 kinase in Drosophila

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