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Digitale Medien

The molecular organisation of a B chromosome tandem repeat sequence from Brachycome dichromosomatica (1996)

Franks, T. K. ; Houben, A. ; Leach, C. R. ; [weitere]

Springer

Chromosoma 105 (1996), S. 223-230

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ISSN: 1432-0886

Quelle: Springer Online Journal Archives 1860-2000

Thema: Biologie , Medizin

Notizen: Abstract. A high copy, tandemly repeated, sequence (Bd49) specific to the B chromosome and located near the centromere in Brachycome dichromosomatica was used to identify lambda genomic clones from DNA of a 3B plant. Only one clone of those analysed was composed entirely of a tandem array of the B-specific repeat unit. In other clones, the Bd49 repeats were linked to, or interspersed with, sequences that are repetitious and distributed elsewhere on the A and B chromosomes. One such repetitious flanking sequence has similarity to retrotransposon sequences and a second is similar to chloroplast DNA sequences. Of the four separate junctions analysed of Bd49-like sequence with flanking sequence, three were associated with the same A/T-rich region in Bd49 and the fourth was close to a 25 bp imperfect dyadic sequence. No novel B-specific sequences were detected within the genomic clones.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1007/s004120050178

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Digitale Medien

Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex (1996)

Houben, A. ; Brandes, A. ; Pich, U. ; [weitere]

Springer

Theoretical and applied genetics 93 (1996), S. 477-484

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ISSN: 1432-2242

Schlagwort(e): Key words Microdissection of plant centromeres ; Subtractive hybridization ; Fluorescence in situ hybridization ; Indirect immunofluorescence ; Immunoadsorption

Quelle: Springer Online Journal Archives 1860-2000

Thema: Biologie

Notizen: Abstract The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are – at least in species with large genomes – intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1007/s001220050305

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3

Digitale Medien

Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex (1996)

Houben, A. ; Brandes, A. ; Pich, U. ; [weitere]

Springer

Theoretical and applied genetics 93 (1996), S. 477-484

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Details

ISSN: 1432-2242

Schlagwort(e): Microdissection of plant centromeres ; Subtractive hybridization ; Fluorescence in situ hybridization ; Indirect immunofluorescence ; Immunoadsorption

Quelle: Springer Online Journal Archives 1860-2000

Thema: Biologie

Notizen: Abstract The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are — at least in species with large genomes — intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1007/BF00417938

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4

Digitale Medien

Immunostaining and interphase arrangement of field bean kinetochores (1995)

Houben, A. ; Guttenbach, M. ; Kreß, W. ; [weitere]

Springer

Chromosome research 3 (1995), S. 27-31

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ISSN: 1573-6849

Schlagwort(e): CREST sera ; interphase kinetochore association ; kinetochore ; nuclear proteins ; Vicia faba

Quelle: Springer Online Journal Archives 1860-2000

Thema: Biologie

Notizen: Abstract More than 100 sera from patients with scleroderma CREST (calcinosis, Raynaud phenomenon, esophageal dismotility, sclerodactyly, telangiectasia) were tested in order to detect antigenic nuclear components of the field beanVicia faba (2n=12). Kinetochores of mitotic chromosomes and prekinetochores of interphase cells from root-tip meristems were specifically labelled via an indirect immunofluorescence procedure by antibodies of one of these sera. In 44% of interphase nuclei in which centromeres could be identified, only half (6) of the number of expected prekinetochores (12) was detected, circumstantially indicating at least transient association of homologous centromeres. Some nuclei showed clustering of centromeres at one pole (Rabl configuration). In metaphase chromosomes, each sister kinetochore contained a fluorescent spot. Western blotting of field bean nuclear proteins revealed four antigenic proteins of 28, 30, 64 and 68 kDa.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1007/BF00711158

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5

Digitale Medien

The acetylation patterns of histones H3 and H4 along Vicia faba chromosomes are different (1998)

Belyaev, N. D. ; Houben, A. ; Baranczewski, P. ; [weitere]

Springer

Chromosome research 6 (1998), S. 59-63

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ISSN: 1573-6849

Schlagwort(e): deacetylase inhibition ; field bean ; heterochromatin ; histone acetylation patterns ; indirect immunofluorescence

Quelle: Springer Online Journal Archives 1860-2000

Thema: Biologie

Notizen: Abstract The acetylation pattern of H3 was studied on field bean chromosomes by means of indirect immunofluorescence using polyclonal antibodies recognizing H3 isoforms acetylated at lysine positions 9/18, 14 and 23. H3 was found to be hypoacetylated at lysine residues 9/18 and 14 within the heterochromatic regions composed of tandem repetitive Fok-I elements. Hyperacetylation of these residues was observed at the nucleolar organizing region (NOR) and in heterochromatic regions composed of repeats other than Fok-I elements. In contrast, H4 was underacetylated (H4.Ac5, 8, 12) or uniformly acetylated (H4.Ac16) at all heterochromatic regions, and acetylated above the average at all four lysines only within the NOR. Acetylation of lysine-23 of H3 was uniform, except for the NOR that showed no fluorescence. Inhibition of deacetylase during and after replication of heterochromatin by trichostatin A had no influence on the acetylation status of H3 but mediated an increase in acetylation of lysines 5, 12 and 16 of H4 above the average in the field bean heterochromatin. Thus, the chromosomal acetylation patterns of H4 and H3 of this species revealed common and divergent features. Whereas the acetylation level of H4 correlates well with the potential transcriptional activity and inversely with the time of replication of defined chromatin domains of Vicia faba, this is not generally true for H3.

Materialart: Digitale Medien

URL: http://dx.doi.org/10.1023/A:1009222609581

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