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1.
Sequence organization and cytological localization of the minor satellite of mouse. 总被引:17,自引:2,他引:15 下载免费PDF全文
A complete 120 bp genomic consensus sequence for the mouse minor satellite has been determined from enriched L929 centromeric sequences. The extensive sequence homology existing between the major and minor satellite suggests an evolutionary relationship. Some sequences flanking the minor satellite has also been identified and they provide insight into centromeric DNA organization. Isotopic in situ hybridization analysis of the minor satellite to mouse L929 and Mus musculus metaphase spreads showed that this repetitive DNA class is localized specifically to centromeres of all chromosomes of the karyotype. With the use of high resolution non-isotopic fluorescence in situ hybridization the minor satellite is further localized to the outer surface of the centromere in a discrete region at or immediately adjacent to the kinetochore. Our cytological data suggests that the minor satellite might play a role in the organization of the kinetochore region rather than, as previously suggested, sites for general anchoring of the genome to the nuclear matrix. 相似文献
2.
David Kipling Helen E. Wilson Arthur R. Mitchell Benjamin A. Taylor Howard J. Cooke 《Chromosoma》1994,103(1):46-55
Cytologically, the centromere is found at the very end of most Mus musculus chromosomes, co-localizing with an array of minor satellite sequences. It is separated from the euchromatin of the long arm by a large domain of heterochromatin, composed in part of arrays of major satellite sequences. We used oligonucleotide probes that specifically detect regions of sequence variation found in certain cloned minor satellite sequences. They detect a limited subset of the minor satellite arrays in the mouse genome, based on both pulsed-field gel electrophoresis and in situ hybridization data, and provide direct molecular genetic markers for individual centromeres in some inbred mouse strains. Array size polymorphisms detected by these probes map to positions consisten with the centromeres of chromosomes 1 and 14 in the BXD recombinant inbred (RI) strains. The genetic distances between these minor satellite arrays and loci on the long arms of chromosomes 1 and 14 are consistent with repression of meiotic recombination in the heterochromatic domains separating them. The existence of chromosome-specific minor satellite sequences implies that the rate of sequence exchange between non-homologous chromosomes relative to the rate between homologous chromosomes is much lower than has previously been postulated. We suggest that the high degree of sequence homogeneity of mouse satellite sequences may instead reflect recent common ancestry. 相似文献
3.
Andrew M. Fisher Lihadh Al-Gazali T. Pramathan Roger Quaife Annette E. Cockwell John C. K. Barber William C. Earnshaw Joyce Axelman Barbara R. Migeon Chris Tyler-Smith 《Chromosoma》1997,106(4):199-206
A de novo dicentric Y;21 (q11.23;p11) translocation chromosome with one of its two centromeres inactive has provided the opportunity
to study the relationship between centromeric inactivation, the organization of alphoid satellite DNA and the distribution
of CENP-C. The proband, a male with minor features of Down’s syndrome, had a major cell line with 45 chromosomes including
a single copy of the translocation chromosome, and a minor one with 46 chromosomes including two copies of the translocation
chromosome and hence effectively trisomic for the long arm of chromosome 21. Centromeric activity as defined by the primary
constriction was variable: in most cells with a single copy of the Y;21 chromosome, the Y centromere was inactive. In the
cells with two copies, one copy had an active Y centromere (chromosome 21 centromere inactive) and the other had an inactive
Y centromere (chromosome 21 centromere active). Three different partial deletions of the Y alphoid array were found in skin
fibroblasts and one of these was also present in blood. Clones of single cell origin from fibroblast cultures were analysed
both for their primary constriction and to characterise their alphoid array. The results indicate that (1) each clone showed
a fixed pattern of centromeric activity; (2) the alphoid array size was stable within a clone; and (3) inactivation of the
Y centromere was associated with both full-sized and deleted alphoid arrays. Selected clones were analysed with antibodies
to CENP-C, and staining was undetectable at both intact and deleted arrays of the inactive Y centromeres. Thus centromeric
inactivation appears to be largely an epigenetic event.
Received: 30 January 1997; in revised form: 3 April 1997 / Accepted: 8 May 1997 相似文献
4.
The centromeres of a genome separate in a sequential, nonrandom manner that is apparently dependent upon the quantity and
quality of pericentric heterochromatin. It is becoming increasingly clear that the biological properties of a centromere depend
upon its physicochemical makeup, such as its tertiary structure, and not necessarily on its particular nucleotide sequence.
To test this idea we altered the physical state of the AT-rich pericentric heterochromatin of mouse with Hoechst 33258 (bis-benzimidazole)
and studied a biological parameter, viz., sequence of separation. We report that an alteration in the physical state of heterochromatin,
i.e., decondensation, is accompanied by aberrations in the pattern of centromere separation. The most dramatic effect seems
to be on chromosomes with large blocks of heterochromatin. Many chromosomes with large blocks of heterochromatin that, in
untreated cells, separate late tend to separate early. Decondensation with Hoechst 33258 does not seem to alter the sequence
of separation of inactive centromeres relative to that of active centromeres. These data indicate that alteration in the physical
parameters of the pericentric heterochromatin might dispose the centromeres to errors. It is likely that this aberration results
from early replication of the pericentric heterochromatin associated with active centromeres.
Received: 24 August 1998; in revised form: 24 August 1998 / Accepted: 28 August 1998 相似文献
5.
Metaphase chromosomes prepared from colcemid-treated mouse L929 cells by non-ionic detergent lysis exhibit distinct heterochromatic centromere regions and associated kinetochores when viewed by whole mount electron microscopy. Deoxyribonuclease I treatment of these chromosomes results in the preferential digestion of the chromosomal arms leaving the centromeric heterochromatin and kinetochores apparently intact. Enrichment in centromere material after DNase I digestion was quantitated by examining the increase in 10,000xg pellets of the 1.691 g/cc satellite DNA relative to main band DNA. This satellite species has been localized at the centromeres of mouse chromosomes by in situ hybridization. From our analysis it was determined that DNase I digestion results in a five to six-fold increase in centromeric material. In contrast to the effect of DNase I, micrococcal nuclease was found to be less selective in its action. Digestion with this enzyme solubilized both chromosome arms and centromeres leaving only a small amount of chromatin and intact kinetochores. 相似文献
6.
Background
The centromere is a specialized locus that mediates chromosome movement during mitosis and meiosis. This chromosomal domain comprises a uniquely packaged form of heterochromatin that acts as a nucleus for the assembly of the kinetochore a trilaminar proteinaceous structure on the surface of each chromatid at the primary constriction. Kinetochores mediate interactions with the spindle fibers of the mitotic apparatus. Centromere protein A (CENP-A) is a histone H3-like protein specifically located to the inner plate of kinetochore at active centromeres. CENP-A works as a component of specialized nucleosomes at centromeres bound to arrays of repeat satellite DNA. 相似文献7.
A method is suggested for isolation of highly purified mouse centromeric heterochromatin. Treatment of mouse liver nuclei with decreasing concentrations of Ca2+ resulted in the gradual unraveling of chromatin in the nucleus and at 0.1 mM Ca2+ electron microscopy revealed several dense particles per nucleus, surrounded by decondensed chromatin. These particles, assumed to represent centromere regions of interphase chromosomes by in situ hybridization with radioactive mouse satellite DNA and by differential staining for centromere heterochromatin, were isolated in preparative amounts and their DNA and protein composition was analyzed. The preparation represented practically pure mouse centromere heterochromatin, since more than 90% of its DNA was satellite DNA. 相似文献
8.
Curvature of mouse satellite DNA and condensation of heterochromatin 总被引:20,自引:0,他引:20
Cloned, sequenced mouse satellite DNA exhibits properties characteristic of molecules that possess a stable curvature. Circularly permuted fragments containing the region predicted to bend were used to map the curvature relative to DNA sequence. The altered mobility of these fragments in polyacrylamide gels is reversed when gels are run in the presence of distamycin A, a drug that binds preferentially to AT-rich DNA. Treatment of living mouse cells with this drug dramatically reduces the condensation of centromeric heterochromatin, the exclusive location of satellite sequences. In situ hybridization of satellite probes to extended chromosomes at the electron microscope level shows that satellite does not comprise a single block but is distributed throughout the centromere region. Based on these experiments, we hypothesize that the structure of mouse satellite DNA is an important feature of centromeric heterochromatin condensation. 相似文献
9.
Caifu Chen Young-Kwon Hong Sara D. Ontiveros Michael Egholm William M. Strauss 《Mammalian genome》1999,10(1):13-18
The satellite repeat structure of the mammalian centromere contains the CENP-B protein binding site. Using the peptide nucleic
acid-fluorescence in situ hybridization (PNA-FISH), we show by direct PNA-DNA binding that all detectable CENP-B sites in
a mammalian genome might have the same sequence. Two species-specific PNA 17-mers, pMm and pMc, were identified from CENP-B
binding sites of Mus musculus and M. caroli, respectively. Fluorescence in situ hybridization confirmed that pMc hybridized to M. caroli centromeres only; however, pMm cross-hybridized to M. musculus and human centromeres. By using a series of CENP-B PNA 17-mers containing 1, 2, 3, 5, and 7 base-pair mismatches to their
DNA counterparts, we further demonstrate that PNA-FISH can discriminate between two CENP-B DNA sequences that differ by a
single base-pair in mouse and human centromeres, suggesting the degree of conservation of CENP-B sequences throughout the
genome. In comparison with DNA oligonucleotides, PNA oligomers demonstrate the higher sequence specificity, improved stability,
reproducibility, and lower background. Therefore, PNA oligomers have significant advantages over DNA oligonucleotide probes
in analyzing microsatellites in a genome.
Received: 16 June 1998 / Accepted: 3 September 1998 相似文献
10.
In situ hybridization of biotin-labeled mouse major satellite DNA clone pMR196 was applied toMus domesticus andMus spretus chromosomes (Chr). The same karyotypes were counterstained with distamycin A-DAPI to identify AT-rich heterochromatin. Chromosomes
from the laboratory mouse, C57BL/6Ros (BL/6;M. domesticus) were uniformly labeled at the centromere except for the Y, while chromosomes from the divergentMus speciesM. spretus showed little or no hybridization. Differences betweenMus species in copy number of the major satellite DNA sequence were used to identify chromosomes ofM. domesticus andM. spretus in their F1 hybrids and to discriminatedomesticus andspretus centromeres in backross progeny. The distribution pattern of heterochromatic regions demonstrated by distamycin A-DAPI counterstaining
was comparable with that of in situ hybridization with pMR196, suggesting that A-T rich heterochromatin inM. domesticus is mainly constructed by the pMR196-related sequence. The in situ technique was used to examine segregation ofdomesticus centromeres in backcross progeny obtained by mating F1 hybrid females withM. domesticus orM. spretus males. The segregation of centromeres did not deviate from the expected among the backcross progeny from C57BL/6Ros males,
whereas chromosomes withM. domesticus centromeres were prone to appear in the progeny from backcross matings byM. spretus males. 相似文献
11.
Heterochromatin and cohesion protection at human centromeres: the final say of a long controversy? 下载免费PDF全文
Jean‐Paul Javerzat 《EMBO reports》2018,19(4)
Mammalian centromeres are embedded within heterochromatin, a specialized chromatin assembled onto repetitive DNA that forms the primary constriction of chromosomes. In early mitosis, the bulk of cohesin dissociates from chromosomes, but a small fraction is spared at the centromere providing the ultimate linker between sister chromatid pairs, essential for their proper attachment to the mitotic spindle. Whether heterochromatin plays a role in the protection of centromere cohesion has long been controversial. In this issue of EMBO Reports, Yi et al show that heterochromatin protein 1 (HP1) isoforms α and γ act redundantly to protect mitotic centromere cohesion through the recruitment of the cohesion protector Haspin 1 . 相似文献
12.
A relationship between the sequence of centromere separation and quantity of pericentromeric constitutive heterochromatin
was studied using bone marrow cells ofMus musculus molossinus and three cell lines, viz., SEWA-Rec 4, brain tumor and L-cells, ofM. m. domesticus origin. The timing of separation of a centromere into two daughter centromeres is related to the quantity of pericentromeric
heterochromatin. In these genomes, having qualitatively uniform DNA in their heterochromatin fraction, the chromosomes with
none or small quantities of heterochromatin separate first. These are followed by those chromosomes which have increasingly
larger quantities of heterochromatin. It appears that one function of repetitive DNA (pericentromeric heterochromatin) is
to regulate the timing of separation of centromeres. 相似文献
13.
Inactive centromeres of stable dicentric chromosomes provide a unique opportunity to examine the resolution of sister chromatid cohesion in mitosis. Here we show for the first time that inactive centromeres are composed of heterochromatin, as defined by the presence of heterochromatin protein HP1(Hs alpha). We then show that both the inner centromere protein (INCENP) and its binding partner Aurora-B/AIM-1 kinase can also be detected at the inactive centromere. Thus, targeting of the chromosomal passengers is not dependent upon the presence of an active centromere/kinetochore. Furthermore, we show that the association of INCENP with the inactive centromere correlates strictly with the state of cohesion between sister chromatids: loss of cohesion is accompanied by loss of detectable INCENP. These results are consistent with recent suggestions that one function of the chromosomal passenger proteins may be to regulate sister chromatid separation in mitosis. 相似文献
14.
Giuliana Giannuzzi Glennis A Logsdon Nicolas Chatron Danny E Miller Julie Reversat Katherine M Munson Kendra Hoekzema Marie-Noëlle Bonnet-Dupeyron Pierre-Antoine Rollat-Farnier Carl A Baker Damien Sanlaville Evan E Eichler Caroline Schluth-Bolard Alexandre Reymond 《Molecular biology and evolution》2021,38(12):5576
Human centromeres are mainly composed of alpha satellite DNA hierarchically organized as higher-order repeats (HORs). Alpha satellite dynamics is shown by sequence homogenization in centromeric arrays and by its transfer to other centromeric locations, for example, during the maturation of new centromeres. We identified during prenatal aneuploidy diagnosis by fluorescent in situ hybridization a de novo insertion of alpha satellite DNA from the centromere of chromosome 18 (D18Z1) into cytoband 15q26. Although bound by CENP-B, this locus did not acquire centromeric functionality as demonstrated by the lack of constriction and the absence of CENP-A binding. The insertion was associated with a 2.8-kbp deletion and likely occurred in the paternal germline. The site was enriched in long terminal repeats and located ∼10 Mbp from the location where a centromere was ancestrally seeded and became inactive in the common ancestor of humans and apes 20–25 million years ago. Long-read mapping to the T2T-CHM13 human genome assembly revealed that the insertion derives from a specific region of chromosome 18 centromeric 12-mer HOR array in which the monomer size follows a regular pattern. The rearrangement did not directly disrupt any gene or predicted regulatory element and did not alter the methylation status of the surrounding region, consistent with the absence of phenotypic consequences in the carrier. This case demonstrates a likely rare but new class of structural variation that we name “alpha satellite insertion.” It also expands our knowledge on alphoid DNA dynamics and conveys the possibility that alphoid arrays can relocate near vestigial centromeric sites. 相似文献
15.
Sequence of centromere separation: differential replication of pericentric heterochromatin in multicentric chromosomes 总被引:2,自引:0,他引:2
The dicentric and multicentric chromosomes in L cells and a brain tumor cell line of mouse display only one site of kinetochore formation associated with the active centromere. The accessory or inactive centromeres show premature separation. These cell lines were treated with 10–6 M 5-bromodeoxyuridine (BrdUrd) followed by anti-BrdUrd antibody to study the pattern of replication of pericentric heterochromatin flanking the active vs inactive centromeres. Regardless of its quantity, heterochromatin around the inactive centromere replicates earlier than that associated with the active centromere. There appears to be a relationship between the timing of separation of a centromere and the timing of replication of pericentric heterochromatin. The premature replication of heterochromatin associated with an inactive centromere may be responsible for its premature separation and, hence, inactivity. 相似文献
16.
The organization of the mouse satellite DNA at centromeres 总被引:2,自引:0,他引:2
The mouse genome contains a major and a minor satellite DNA family of repetitive DNA sequences. The use of 5-azacytidine has allowed us to demonstrate that these satellite DNAs are organized in two separate domains at the centromeres of mouse chromosomes. The minor satellite is closer to the short arms of the acrocentric chromosomes than the major satellite. The major satellite is farther away, flanking the minor satellite and adjacent to the euchromatic long arm of each mouse chromosome. At the level of resolution afforded by the in situ hybridization technique it would appear that the organization of the centromeric domain of the mouse is similar to that in man. That is, both contain two repetitive DNA sequence families arranged in major blocks. 相似文献
17.
Reactivation of an Inactive Centromere Reveals Epigenetic and Structural Components for Centromere Specification in Maize 总被引:1,自引:0,他引:1
Stable maize (Zea mays) chromosomes were recovered from an unstable dicentric containing large and small versions of the B chromosome centromere. In the stable chromosome, the smaller centromere had become inactivated. This inactive centromere can be inherited from one generation to the next attached to the active version and loses all known cytological and molecular properties of active centromeres. When separated from the active centromere by intrachromosomal recombination, the inactive centromere can be reactivated. The reactivated centromere regains the molecular attributes of activity in anaphase I of meiosis. When two copies of the dicentric chromosome with one active and one inactive centromere are present, homologous chromosome pairing reduces the frequency of intrachromosomal recombination and thus decreases, but does not eliminate, the reactivation of inactive centromeres. These findings indicate an epigenetic component to centromere specification in that centromere inactivation can be directed by joining two centromeres in opposition. These findings also indicate a structural aspect to centromere specification revealed by the gain of activity at the site of the previously inactive sequences. 相似文献
18.
Hasson D Alonso A Cheung F Tepperberg JH Papenhausen PR Engelen JJ Warburton PE 《Chromosoma》2011,120(6):621-632
Endogenous human centromeres form on megabase-sized arrays of tandemly repeated alpha satellite DNA. Human neocentromeres
form epigenetically at ectopic sites devoid of alpha satellite DNA and permit analysis of centromeric DNA and chromatin organization.
In this study, we present molecular cytogenetic and CENP-A chromatin immunoprecipitation (ChIP) on CHIP analyses of two neocentromeres
that have formed in chromosome band 8q21 each with a unique DNA and CENP-A chromatin configuration. The first neocentromere
was found on a neodicentric chromosome 8 with an inactivated endogenous centromere, where the centromeric activity and CENP-A
domain were repositioned to band 8q21 on a large tandemly repeated DNA. This is the first example of a neocentromere forming
on repetitive DNA, as all other mapped neocentromeres have formed on single copy DNA. Quantitative fluorescent in situ hybridization
(FISH) analysis showed a 60% reduction in the alpha satellite array size at the inactive centromere compared to the active
centromere on the normal chromosome 8. This neodicentric chromosome may provide insight into centromere inactivation and the
role of tandem DNA in centromere structure. The second neocentromere was found on a neocentric ring chromosome that contained
the 8q21 tandemly repeated DNA, although the neocentromere was localized to a different genomic region. Interestingly, this
neocentromere is composed of two distinct CENP-A domains in bands 8q21 and 8q24, which are brought into closer proximity on
the ring chromosome. This neocentromere suggests that chromosomal rearrangement and DNA breakage may be involved in neocentromere
formation. These novel examples provide insight into the formation and structure of human neocentromeres. 相似文献
19.
AluI and BstNI restriction endonucleases were used to study cytological and biochemical effects on centromere DNA in fixed mouse chromosomes. These enzymes were employed, as it is known that AluI is incapable of attacking major satellite DNA, contrary to BstNI that is known to cut this DNA fraction into monomers of 234 bp. After digestion in situ, electrophoretic analysis was carried out to characterize the DNA purified (1) from the material remaining on the chromosomes and (2) from the material solubilized from chromosomes. The DNA was then transferred to a nylon filter and 32P-labelled major satellite DNA was used as a probe for hybridization experiments. Other preparations were simply stained with Giemsa after digestion in situ with AluI and BstNI. Our results show that although restriction endonuclease cleavage primarily depends on DNA base sequence, this factor is not always sufficient to explain nuclease-induced cytological effects. In fact, the structural organization of peculiar regions such as the centromeres of mouse chromosomes might affect cleavage efficiency when restriction enzyme digestion is performed in situ.M.L. Pardue 相似文献
20.
Plant centromeres are generally composed of tandem arrays of simple repeats that form a complex chromosome locus where the kinetochore forms and microtubules attach during mitosis and meiosis. Each chromosome has one centromere region, which is essential for accurate division of the genetic material. Recently, chromosomes containing two centromere regions (called dicentric chromosomes) have been found in maize and wheat. Interestingly, some dicentric chromosomes are stable because only one centromere is active and the other one is inactivated. Because such arrays maintain their typical structure for both active and inactive centromeres, the specification of centromere activity has an epigenetic component independent of the DNA sequence. Under some circumstances, the inactive centromeres may recover centromere function, which is called centromere reactivation. Recent studies have highlighted the important changes, such as DNA methylation and histone modification, that occur during centromere inactivation and reactivation. 相似文献