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  • 1
    Electronic Resource
    Electronic Resource
    Enhanced processivity of nuclear matrix bound DNA polymerase .alpha. from regenerating rat liver (1987)
    s.l. : American Chemical Society
    Biochemistry 26 (1987), S. 5710-5718 
    Details
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/bi00392a020
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  • 2
    Electronic Resource
    Electronic Resource
    An immunohistochemical study of protein kinase C distribution in fetal mouse vertebral column (1994)
    Springer
    Anatomy and embryology 190 (1994), S. 47-54 
    Details
    ISSN: 1432-0568
    Keywords: Protein kinase C ; Isozymes ; Immunolocalization ; Vertebral column ; Mouse development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using polyclonal antibodies we have studied the distribution of protein kinase C in fetal mouse low thoracic vertebrae. By means of a pan protein kinase C antiserum recognizing the catalytic domain of the enzyme, we show that protein kinase C is markedly expressed in chondrocytes before birth. The enzyme seems to be very abundant in the more mature cells that are close to ossification centres as well as the periphery of the intervertebral disc, although it can also be detected in chondrocytes. In order to establish which protein kinase C isoenzyme(s) the chondrocytes produce, we employed polyclonal isoenzyme-specific antisera developed against three calcium-dependent isoforms (α, β, γ) and three calcium-independent isoforms (gd, ɛ, ζ). Secondary antibody conjugated to alkaline phosphatase revealed that chondrocytes markedly express the β-isoform. Cells were also weakly stained by the anti-ɛ serum. The immunostaining was completely abolished by pre-incubating primary antibodies with the peptide antigens to which they were raised. These results suggest that protein kinase C (and particularly the β isoform) could play an important role in mouse fetal chondrogenesis of the vertebral column.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00185845
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  • 3
    Electronic Resource
    Electronic Resource
    Nuclear localization and signalling activity of phosphoinositidase Cβ in Swiss 3T3 cells (1992)
    [s.l.] : Nature Publishing Group
    Nature 358 (1992), S. 242-245 
    Details
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] An analysis of phosphoinositidase C (PIC) activities in the nucleus and cytoplasm of Swiss 3T3 cells using [3H]PtdInsP and [3H]PtdInsP2 as substrates is shown in Fig. 1. Both nuclear and cytoplasmic activities require calcium because the addition of EGTA completely abolishes breakdown. ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/358242a0
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    Paper (German National Licenses)
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  • 4
    Electronic Resource
    Electronic Resource
    Subnuclear localization of S/MAR-binding proteins is differently affected by in vitro stabilization with heat or Cu2+ (1997)
    Springer
    Chromosoma 106 (1997), S. 81-93 
    Details
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The nuclear matrix, a proteinaceous entity thought to be a scaffolding structure that determines the higher order organization of eukaryotic chromatin, is usually prepared from intact nuclei by a series of extraction steps. In most cell types investigated, the nuclear matrix does not spontaneously resist these extractions, but must rather be stabilized before the application of extracting agents such as high salt solutions or lithium diiodosalicylate. We have examined the effect of two widely used stabilization procedures on the localization of nuclear matrix proteins. Four individual polypeptides were studied, all of which are scaffold or matrix-associated region (S/MAR)-binding proteins: SATB1, SAF-A/hnRNP-U, NuMA , and topoisomerase II α. Nuclei were isolated from K562 human erythroleukemia cells in a buffer containing spermine, spermidine, KCl and EDTA, and the nuclear matrix or scaffold was obtained by extraction with lithium diiodosalicylate after stabilization by heat treatment (37° or 42°C) or incubation with Cu2+ ions. When the localization of individual proteins was determined by immunofluorescent staining and confocal scanning laser microscopy, markedly different consequences of the two stabilization strategies became evident, ranging from a total maintenance of the localization (NuMA and topoisomerase II α) to a marked redistribution (SATB1 and SAF-A/hnRNP-U). Our results seem to indicate that a reevaluation of stabilization protocols employed for the preparation of the nuclear matrix is desirable, especially by performing morphological controls.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004120050227
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  • 5
    Electronic Resource
    Electronic Resource
    Protein kinase C isoenzymes in mouse Harderian gland. Differential expression of the α- and ε-isoforms during pregnancy (1995)
    Springer
    Histochemistry and cell biology 103 (1995), S. 255-262 
    Details
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Protein kinase C (PKC) is known to be involved in the regulation of exocytosis in different cell lines and tissues. Experiments were designed to determine whether the Harderian gland of CD-1 mouse produces PKC isoenzymes and whether the expression of the isoforms changes during pregnancy. The presence of the isoenzymes was assessed by immunoblotting experiments using extract of total Harderian gland and polyclonal antisera specific for nine different PKC isoforms. Antisera giving a positive staining on Western blots were subsequently used for immunohistochemical investigation using a secondary antibody conjugated to alkaline phosphatase. Immunoblotting experiments revealed that the Harderian gland from female mouse expresses PKC isoforms-α,-ε,-ζ and-η. These isoforms were also detected in the Harderian gland from 13-day pregnant mouse; however, striking quantitative changes were seen concerning the α- and ε-isoforms. The 80-kDa native form of PKC-α almost doubled in the pregnant mouse in comparison with normal female mouse whereas the amount of 50-kDa catalytic domain did not change. Protein kinase C-ε appeared as a 92- to 93-kDa form and a 67-kDa form. While the 92- to 93-kDa protein was expressed to a similar extent in both types of mouse, the 67-kDa form was more abundant inthe Harderian gland from normal female mouse. These data were corroborated by immunohistochemical experiments and showing a diffuse and granular staining of the adenomeres. These observations demonstrate for the first time (to our knowledge) that the mouse Harderian gland produces several PKC isoenzymes that could be involved in the regulation of exocytosis and/or other functions. Moreover, the expression of the α- and ε-isoforms could be regulated by sexual hormones, as suggested by the differential abundance of these two proteins in the gland of pregnant mouse compared with normal female mouse.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01457409
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  • 6
    Electronic Resource
    Electronic Resource
    Distribution of the extended family of protein kinase C isoenzymes in fetal organs of mice: an immunohistochemical study (1995)
    Springer
    Cell & tissue research 280 (1995), S. 617-625 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Protein kinase C ; Isozymes- Immunolocalization ; Immunoblotting ; Fetal organs ; Mouse (CD-1)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Using isoenzyme-specific antisera, we have studied the distribution of protein kinase C isoforms in fetal mouse organs at the developmental age of 17 days. Two different sets of antibodies, produced by different manufacturers, were employed in this study. The specificity of the antisera was tested by immunoblotting experiments using whole fetal mouse extracts. Immunohistochemistry was carried out by means of an alkaline phosphatase-conjugated secondary antibody. Analysis of fetal mouse longitudinal cryostat sections stained with the antibodies demonstrated a distinct distribution of protein kinase C isoforms in the tissues. Protein kinase C-α and C-βI were present in all tissues examined, whereas the C-βII isoform was absent in the lung and the liver. Protein kinase C-γ was identified in brain, spinal ganglia, and adrenal gland. The C-ɛ isoenzyme was abundantly expressed in spinal ganglia and in the smooth muscle cells of the bronchial wall. Antisera to C-ζ and C-η isoforms heavily stained liver, kidney, and spinal ganglia, whereas the C-θ isozyme was mainly detected in brain, stomach and kidney. Thus, protein kinase C-α, C-βI, C-βII, C-ζ , C-η and C-θ were the isoforms present in many of the organs investigated. The two sets of antibodies gave slightly different results that might be ascribed to the different epitopes recognized by the antisera. One set of antisera was employed to investigate the distribution of the isoforms in selected organs from an earlier developmental age (15 days) and from adult animals. Both qualitative and quantitative differences were seen in comparison with the same organs from a 17-day fetus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318364
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  • 7
    Electronic Resource
    Electronic Resource
    Distribution of the extended family of protein kinase C isoenzymes in fetal organs of mice: an immunohistochemical study (1995)
    Springer
    Cell & tissue research 280 (1995), S. 617-625 
    Details
    ISSN: 1432-0878
    Keywords: Protein kinase C ; Isozymes ; Immunolocalization ; Immunoblotting ; Fetal organs ; Mouse (CD-1)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using isoenzyme-specific antisera, we have studied the distribution of protein kinase C isoforms in fetal mouse organs at the developmental age of 17 days. Two different sets of antibodies, produced by different manufacturers, were employed in this study. The specificity of the antisera was tested by immunoblotting experiments using whole fetal mouse extracts. Immunohistochemistry was carried out by means of an alkaline phosphatase-conjugated secondary antibody. Analysis of fetal mouse longitudinal cryostat sections stained with the antibodies demonstrated a distinct distribution of protein kinase C isoforms in the tissues. Protein kinase C-α and C-βI were present in all tissues examined, whereas the C-βII isoform was absent in the lung and the liver. Protein kinase C-γ was identified in brain, spinal ganglia, and adrenal gland. The C-ε isoenzyme was abundantly expressed in spinal ganglia and in the smooth muscle cells of the bronchial wall. Antisera to C-ζ and C-η isoforms heavily stained liver, kidney, and spinal ganglia, whereas the C-θ isozyme was mainly detected in brain, stomach and kidney. Thus, protein kinase C-α, C-βI, C-βII, C-ζ, C-η and C-θ were the isoforms present in many of the organs investigated. The two sets of antibodies gave slightly different results that might be ascribed to the different epitopes recognized by the antisera. One set of antisera was employed to investigate the distribution of the isoforms in selected organs from an earlier developmental age (15 days) and from adult animals. Both qualitative and quantitative differences were seen in comparison with the same organs from a 17-day fetus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00318364
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    Paper (German National Licenses)
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  • 8
    Electronic Resource
    Electronic Resource
    On the association of DNA primase activity with the nuclear matrix in HeLa S3 cells (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Cell Biochemistry and Function 11 (1993), S. 287-290 
    Details
    ISSN: 0263-6484
    Keywords: DNA primase ; nuclear matrix ; nuclear isolation ; heat exposure ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have reinvestigated the association of DNA primase activity with the nuclear matrix prepared from exponentially growing HeLa S3 cells. We have found that 25-30 per cent of the nuclear primase activity resists extraction with 2 M NaCl and digestion with Dnase I. Unlike previous investigations, done with the same cell line, the results showed that nuclear matrix-bound DNA primase activity represented less than 10 per cent of the total cell activity. Association of high levels of primase activity with the nuclear matrix was strictly dependent on a 37°C incubation of isolated nuclei prior to subfractionation. Evidence was obtained that the method used for preparing nuclei can have a dramatic effect on the amount of primase activity which is recovered both in the postnuclear supernatant and in isolated nuclei, thus seriously affecting the interpretation of the results about the quantity of DNA primase activity bound to the nuclear matrix.
    Additional Material: 3 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cbf.290110410
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  • 9
    Electronic Resource
    Electronic Resource
    No discrete complexes containing DNA polymerase α activity can be solubilized from the heat-stabilized nuclear matrix prepared from HeLa S3 cells (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Cell Biochemistry and Function 12 (1994), S. 37-44 
    Details
    ISSN: 0263-6484
    Keywords: Nuclear matrix ; DNA polymerase α ; soluble complexes ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Most of the DNA polymerase α activity, bound to the heat-stabilized nuclear matrix prepared from HeLa S3 cells, was released as a matrix extract by sonication. When the extract was centrifuged in a 5-20 per cent linear sucrose gradient no definite peaks of activity could be identified. Most of the activity sedimented to the bottom of the tube under all the conditions tested, whilst the remaining activity was associated with matrix fragments of various and irregular size. No 10 S complexes, containing polymerase activity, were seen after incubation of the extract for 16 h before centrifugation. Other solubilization procedures (i.e. treatment of the matrix with chelating agents, high pH associated with reducing agents, ionic and nonionic detergents) failed to produce release of matrix-bound DNA polymerase α activity. In contrast, we released 10 S complexes, containing polymerase activity, from the matrix prepared from nuclei not exposed to heat. We conclude that a 37°C incubation of isolated nuclei before extraction with 2 M NaCl and DNase I digestion causes DNA polymerase α to bind to the nuclear matrix in a form that cannot subsequently be released as discrete components, at variance with previous results obtained with the matrix prepared from regenerating rat liver.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cbf.290120106
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  • 10
    Electronic Resource
    Electronic Resource
    Catalytic properties of DNA polymerase α activity associated with the heat-stabilized nuclear matrix prepared from HeLa S3 cells (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Cell Biochemistry and Function 12 (1994), S. 129-135 
    Details
    ISSN: 0263-6484
    Keywords: Nuclear matrix ; DNA polymerase α ; processivity ; activity ; heat stabilization ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have investigated whether or not ATP or other nucleoside di- and trisphosphates (including some nonhydrolysable ATP analogues) can stimulate the activity and/or the processivity of DNA polymerase α associated with the nuclear matrix obtained from HeLa S3 cell nuclei that had been stabilized at 37°C prior to subfractionation, as has been reported previously for DNA polymerase α bound to the nuclear matrix prepared from 22-h regenerating rat liver. We have found that HeLa cell matrix-associated DNA polymerase α activity could not be stimulated at all by ATP or other nucleotides, a behaviour which was shared also by DNA polymerase α activity that solubilizes from cells during the isolation of nuclei and that is thought to be a form of the enzyme not actively engaged in DNA replication. Moreover, the processivity of matrix-bound DNA polymerase α activity was low (〈 10 nucleotides). These results were obtained with the matrix prepared with either 2M NaCl or 0·25 M (NH4)2SO4 and led us to consider that a 37° incubation of isolated nuclei renders resistant to high-salt extraction a form of DNA polymerase α which is unlikely to be involved in DNA replication in vivo.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cbf.290120208
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