Localization of 5-methylcytosine in human metaphase chromosomes by immunoelectron microscopy

Lubit, Beverly W.; Duc, Pham Tuan; Miller, Orlando J.; Erlanger, Bernard F.

doi:10.1016/0092-8674(76)90032-5

Cited by 58 publications

(13 citation statements)

References 28 publications

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“…In human metaphase chromosomes there is also complete correlation between the distamycin A/DAPI positive regions, the presence of 5-methylcytosine as detected by immunochemical techniques (Miller et al 1974;Lubit et al 1976), and undercondensation brought about by treatment with 5-azacytidine (Schmid et al 1983). Since almost all 5-methylcytosine in mammalian DNA occurs in the dinucleotide sequence 5' CpG 3' (Doskocil and Sorm 1962) it may be more than coincidental that the single base pair change in the DI5Z1 consensus sequence that results in both HinfI and TaqI sites, also introduces CpG dinucleotides into a sequence otherwise devoid of them.…”

Section: Discussionmentioning

confidence: 93%

Organization of a repetitive human 1.8 kb KpnI sequence localized in the heterochromatin of chromosome 15

et al. 1985

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We have isolated a repetitive 1.8 kb KpnI DNA sequence which is amplified in the homogeneously staining regions of a human melanoma cell line. Under low stringency conditions this sequence (D15Z1) hybridized in situ to the centromeric heterochromatin of chromosomes 1, 9, 15p, 16, and distal Yq as well as to the short arms of the other acrocentric chromosomes. Under conditions of high stringency, labelling was predominantly on the short arm of chromosome 15. D15Z1 was shown to be present at approximately 3,000 copies per haploid genome and organized in long tandem arrays showing restriction site heterogeneity. Sequences homologous to D15Z1 were highly enriched in the less dense shoulder region of a Ag+-Cs2SO4 gradient. Analysis of D15Z1 indicated that this sequence is composed of tandemly arranged imperfect repeats of the consensus 5' AATGG 3' similar to previously identified satellite III sequences. Digestion of D15Z1 with HinfI resulted in a series of restriction fragments making up a subset of the HinfI ladder components of satellites III and IV. These data suggest that D15Z1 represents a chromosome 15 specific domain of human satellites III or IV and that it makes up the major fraction of the heterochromatin of this chromosome. Possible relationships between this sequence and the cytochemical staining properties of human chromosomes with distamycin A/DAPI, D280/170, and antiserum to 5-methylcytosine are discussed.

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Section: Discussionmentioning

confidence: 93%

Organization of a repetitive human 1.8 kb KpnI sequence localized in the heterochromatin of chromosome 15

et al. 1985

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“…Secondly, the plastic film used to transfer the mitotic figures must be readily removable from the glass slide. Hydrofluoric acid, which was used by Lubit et al (1976) and Ruzicka (1977) to detach chromosome-carrying carbon film from glass slides, is so effective that no difficulty should be encountered in removing the small plastic disc. The chemical does not seem to affect the chro mosome morphology.…”

Section: Resultsmentioning

confidence: 99%

Easy transfer of selected mitoses from light to electron microscopy

Messier¹,

Jean²,

Richer³

1986

Cytogenet Genome Res

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“…It is well known that modification of DAPI banding on chromosomes by prestaining with the DNA-binding oligopeptide antibiotic distamycin A reveals a specific subset of AT base pair-rich heterochromatin [Sumner, 1990]. Schweizer et al [1978] noted that the distamycin A/DAPI-positive heterochromatic regions in human chromosomes correspond exactly to those sites enriched in 5-methylcytosine Lubit et al, 1976;Schnedl et al, 1976]. However, this correlation cannot be generalized because, for example, the highly repetitive satellite DNA of the mouse (Mus musculus) is highly methylated, but not distamycin A/ DAPI-positive [Schnedl et al, 1980].…”

Section: -Mec-rich Heterochromatinmentioning

confidence: 99%

Chromosome Banding in Amphibia. XXXIII. Demonstration of 5-Methylcytosine-Rich Heterochromatin in Anura

Schmid¹,

Steinlein²

2016

Cytogenet Genome Res

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An experimental approach using monoclonal anti-5-methylcytosine (5-MeC) antibodies and indirect immunofluorescence was elaborated for detecting 5-MeC-rich chromosome regions in anuran chromosomes. This technique was applied to mitotic metaphases of 6 neotropical frog species belonging to 6 genera and 4 families. The hypermethylation patterns were compared with a variety of banding patterns obtained by conventional banding techniques. The hypermethylated DNA sequences are species-specific and located exclusively in constitutive heterochromatin. They are found in centromeric, pericentromeric, telomeric, and interstitial positions of the chromosomes and adjacent to nucleolus organizer regions. 5-MeC-rich DNA sequences can be embedded both in AT- and GC-rich repetitive DNA. The experimental parameters that have major influence on the reproducibility and quality of the anti-5-MeC antibody labeling are discussed.

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Localization of 5-methylcytosine in human metaphase chromosomes by immunoelectron microscopy

Cited by 58 publications

References 28 publications

Organization of a repetitive human 1.8 kb KpnI sequence localized in the heterochromatin of chromosome 15

Organization of a repetitive human 1.8 kb KpnI sequence localized in the heterochromatin of chromosome 15

Easy transfer of selected mitoses from light to electron microscopy

Chromosome Banding in Amphibia. XXXIII. Demonstration of 5-Methylcytosine-Rich Heterochromatin in Anura

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