Dodeca satellite: a conserved G+C-rich satellite from the centromeric heterochromatin of Drosophila melanogaster.

Abad, José Pascual; Carmena, Mar; Baars, Sigrid; Saunders, Robert D. C.; Glover, David M.; Ludeña, P.; Sentís, C.; Tyler‐Smith, Chris; Villasanté, Alfredo

doi:10.1073/pnas.89.10.4663

Cited by 103 publications

(92 citation statements)

References 18 publications

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“…Centromeric sequences do not seem to be broadly conserved throughout evolution, although examples of conserved centromeric satellite DNAs in related species are abundant. Dodeca satellite is present in the centromeric region of chromosome 3 of D. melanogaster and in the centromeric region of chromosomes 2 and 3 of Drosophila simulans and Drosophila mauritiana (10,19). Centromeric nucleosomes contain the centromere-specific histone H3 variant CENP-A, CID in Drosophila (57,58).…”

Section: Immunocytological Analysis Of Chromosomal Proteins Within Thementioning

confidence: 99%

“…However, their precise localization relative to other sequenced portions of the genome remains unknown. In situ hybridization on mitotic chromosomes has been successfully used to map bacterial artificial chromosomes (BACs) (4, 9, 13-16) as well as highly repeated DNA sequences (9,(17)(18)(19)(20). Unfortunately, resolution of this approach remains low (Ϸ1 Mb) because of the small linear size of mitotic chromosome.…”

mentioning

confidence: 99%

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High-resolution analysis of Drosophila heterochromatin organization using SuUR Su(var)3-9 double mutants

Andreyeva

Колесникова

Demakova

et al. 2007

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

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The structural and functional analyses of heterochromatin are essential to understanding how heterochromatic genes are regulated and how centromeric chromatin is formed. Because the repetitive nature of heterochromatin hampers its genome analysis, new approaches need to be developed. Here, we describe how, in double mutants for Su(var)3-9 and SuUR genes encoding two structural proteins of heterochromatin, new banded heterochromatic segments appear in all polytene chromosomes due to the strong suppression of under-replication in pericentric regions. FISH on salivary gland polytene chromosomes from these double mutant larvae allows high resolution of heterochromatin mapping. In addition, immunostaining experiments with a set of antibodies against euchromatic and heterochromatic proteins reveal their unusual combinations in the newly appeared segments: binding patterns for HP1 and HP2 are coincident, but both are distinct from H3diMetK9 and H4triMetK20. In several regions, partial overlapping staining is observed for the proteins characteristic of active chromatin RNA Pol II, H3triMetK4, Z4, and JIL1, the boundary protein BEAF, and the heterochromatin-enriched proteins HP1, HP2, and SU(VAR)3-7. The exact cytological position of the centromere of chromosome 3 was visualized on salivary gland polytene chromosomes by using the centromeric dodeca satellite and the centromeric protein CID. This region is enriched in H3diMetK9 and H4triMetK20 but is devoid of other proteins analyzed.Drosophila melanogaster ͉ genome analysis

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Section: Immunocytological Analysis Of Chromosomal Proteins Within Thementioning

confidence: 99%

mentioning

confidence: 99%

High-resolution analysis of Drosophila heterochromatin organization using SuUR Su(var)3-9 double mutants

Andreyeva

Колесникова

Demakova

et al. 2007

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

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“…Since the chromocenter represents mostly repetitive sequences within constitutive heterochromatin, we tested if the replication timing of centromeric repeats on chromosome 3 is advanced after HP1 RNAi. To this end, we used primers specific for the dodeca repeat sequence located in pericentric heterochromatin of chromosome 3 (Abad et al 1992). In untreated cells, we detected the highest enrichment of replicating DNA in S4, representing the latest S-phase fraction (Fig.…”

Section: Advanced Replication Of Centromeric Repeats After Hp1 Knockdownmentioning

confidence: 99%

Heterochromatin protein 1 (HP1) modulates replication timing of the Drosophila genome

Schwaiger¹,

Kohler²,

Oakeley³

et al. 2010

Genome Res.

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The replication of a chromosomal region during S phase can be highly dynamic between cell types that differ in transcriptome and epigenome. Early replication timing has been positively correlated with several histone modifications that occur at active genes, while repressive histone modifications mark late replicating regions. This raises the question if chromatin modulates the initiating events of replication. To gain insights into this question, we have studied the function of heterochromatin protein 1 (HP1), which is a reader of repressive methylation at histone H3 lysine 9, in genome-wide organization of replication. Cells with reduced levels of HP1 show an advanced replication timing of centromeric repeats in agreement with the model that repressive chromatin mediates the very late replication of large clusters of constitutive heterochromatin. Surprisingly, however, regions with high levels of interspersed repeats on the chromosomal arms, in particular on chromosome 4 and in pericentromeric regions of chromosome 2, behave differently. Here, loss of HP1 results in delayed replication. The fact that these regions are bound by HP1 suggests a direct effect. Thus while HP1 mediates very late replication of centromeric DNA, it is also required for early replication of euchromatic regions with high levels of repeats. This observation of opposing functions of HP1 suggests a model where HP1-mediated repeat inactivation or replication complex loading on the chromosome arms is required for proper activation of origins of replication that fire early. At the same time, HP1-mediated repression at constitutive heterochromatin is required to ensure replication of centromeric repeats at the end of S phase.

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“…Various heterochromatic probes were used: X-chromosome 359 satellite chromosome 2 AACAC satellite and chromosome 3 dodecasatellite (Abad et al, 1992). Euchromatic BAC clones were obtained from the MRC UK HGMP resource centre, Cambridge, UK (BACH47E07, 10A-10A -X chromosome-; BACHN09, 30C5-30D -chromosome 2L-; BACH47E02, 50A1-50A5 -chromosome 2R-; BACH47K04, 71A-71A -chromosome 3L-; BACN01B01, 90C7-90C10 -chromosome 3R-.…”

Section: Fluorescent In Situ Hybridisation (Fish)mentioning

confidence: 99%

RNAi analysis reveals an unexpected role for topoisomerase II in chromosome arm congression to a metaphase plate

Chang

Goulding

Earnshaw

et al. 2003

Journal of Cell Science

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DNA topoisomerase II (Topo II) is a major component of mitotic chromosomes and an important drug target in cancer chemotherapy, however, its role in chromosome structure and dynamics remains controversial. We have used RNAi to deplete Topo II in Drosophila S2 cells in order to carry out a detailed functional analysis of the role of the protein during mitosis. We find that Topo II is not required for the assembly of a functional kinetochore or the targeting of chromosomal passenger proteins, nonetheless, it is essential for anaphase sister chromatid separation. In response to a long-running controversy, we show that Topo II does have some role in mitotic chromatin condensation. Chromosomes formed in its absence have a 2.5-fold decrease in the level of chromatin compaction, and are morphologically abnormal. However, it is clear that the overall programme of mitotic chromosome condensation can proceed without Topo II. Surprisingly, in metaphase cells depleted of Topo II, one or more chromosome arms frequently stretch out from the metaphase plate to the vicinity of the spindle pole. This is not kinetochore-based movement, as the centromere of the affected chromosome is located on the plate. This observation raises the possibility that further unexpected functions for Topo II may remain to be discovered.

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Dodeca satellite: a conserved G+C-rich satellite from the centromeric heterochromatin of Drosophila melanogaster.

Cited by 103 publications

References 18 publications

High-resolution analysis of Drosophila heterochromatin organization using SuUR Su(var)3-9 double mutants

High-resolution analysis of Drosophila heterochromatin organization using SuUR Su(var)3-9 double mutants

Heterochromatin protein 1 (HP1) modulates replication timing of the Drosophila genome

RNAi analysis reveals an unexpected role for topoisomerase II in chromosome arm congression to a metaphase plate

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