DNA Sequence-Specific Location of Covalent DNA-Polypeptide Complexes in Eukaryotic Genomes

Werner, Dieter; Pfütz, Marita

doi:10.1007/978-1-4613-0667-2_2

Cited by 1 publication

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References 13 publications

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“…Distinct proteins designated as the most tightly bound proteins in eukaryotic DNA (54-68 kDa) that are not released by alkali, SDS, moderate protease treatment, have been identified in a variety of cell types upon extensive DNase I degradation of DNA purified by standard procedures (Werner et al, 1981, Neuer et a!., 1983. The site-specific location in eukaryotic genomes (Werner and Neuer-Nitsche, 1989) and association with the nuclear matrix (Werner and Pfutz, 1990) of these tight (and/or covalent) DNApolypeptide complexes has been shown. The Mab 2A8 epitope persisted in the fraction of such residual nuclease-and alkali-resistant polypeptides isolated from diverse cell types, fibroblast cells, sperm, thymus, and is apparently a common structurally conserved motif of a DNA tight binding protein.…”

Section: Discussionmentioning

confidence: 99%

Redistribution of nuclear envelope associated antigen during the mitotic cycle.

Batova

Kyurkchiev²,

Russev³

et al. 1995

Cell Biology International

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Murine hybridomas were generated to DNA/tight binding proteins complex isolated from the residual nuclear structure following a procedure analogous to that yielding "empty" shells of nuclear envelope. A monoclonal antibody designated 2A8 was selected because of its differential immunostaining of mitotic cells of a synchronized mouse fibroblast cell culture L-929. The target antigen was rendered insoluble by a sequence of extractions of isolated nuclei of diverse cell types with detergents, urea, DNase I and alkali thus reproducing some solubility properties of proteins constituting an operationally defined residual nuclear matrix. The cognate polypeptide was localized on a subset of proteins of M(r) 58-65 kDa, 70 kDa in isolated fibroblast nuclear matrices. The functional implication of the antigen in mitosis-related disassembly-assembly process of the nuclear matrix/envelope was detected. At prophase the antibody decorated the nuclear periphery and nuclear envelope fixed inward filaments. A fibrous network of cytoplasmic localization was stained in metaphase. At anaphase the antigen was dispositioned into peripheral fibrogranular clusters of polar orientation predominantly on one side of the nucleus. Proceeding to telophase a spreading fluorescence was manifested over the entire contour of the nuclear periphery to delineate the reforming nucleus. By immunogold electron microscopy of interphase cells the antigen was identified as evenly distributed in chromatin and interchromatin regions. At initiation of chromosome condensation in mitosis the label was detected predominantly in the chromosomal area.

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