scholarly journals Immunoglobulins anti-endonuclease 32 kDa from Cucurbita pepo var patissonina affect process of DNA synthesis.

1995 ◽  
Vol 42 (2) ◽  
pp. 177-182
Author(s):  
R Rzepecki ◽  
J Szopa
Keyword(s):  
Dna Synthesis ◽  
Nuclear Matrix ◽  
Protein Complex ◽  
Cucurbita Pepo ◽  
Dna Fragments ◽  
Cell Nuclei ◽  
Structural Organisation ◽  
Low Concentrations

Immunoglobulins anti-endonuclease 32 kDa inhibit DNA synthesis. We observed that low concentrations of IgGs (about 50 micrograms IgG per 1 x 10(6) cell nuclei) temporary inhibit DNA synthesis. This inhibition concerns only the synthesis of DNA bound to the nuclear matrix (associated with isolated nuclear matrix). Preincubation of cell nuclei of White bush with IgG generates longer DNA fragments than in controls. Involvement of the 32 kDa endonuclease or an endonuclease-65 kDa protein complex from the nuclear matrix in replication or structural organisation of replication is considered.

Use of Antibody to aid Visualization of Protein at the Termini of Bacteriophage Ø29 DNA

1980 ◽  
Vol 38 ◽  
pp. 540-541
Author(s):  
Dwight Anderson ◽  
Charlene Peterson ◽  
Gursaran Notani ◽  
Bernard Reilly
Keyword(s):  
Electron Microscopy ◽  
Bacillus Subtilis ◽  
Dna Synthesis ◽  
Restriction Endonuclease ◽  
Protein Complex ◽  
Protein Product ◽  
Viral Dna ◽  
Small Proteins ◽  
Antibody Labeling ◽  
Subtilis Bacteriophage

The protein product of cistron 3 of Bacillus subtilis bacteriophage Ø29 is essential for viral DNA synthesis and is covalently bound to the 5’-termini of the Ø29 DNA. When the DNA-protein complex is cleaved with a restriction endonuclease, the protein is bound to the two terminal fragments. The 28,000 dalton protein can be visualized by electron microscopy as a small dot and often is seen only when two ends are in apposition as in multimers or in glutaraldehyde-fixed aggregates. We sought to improve the visibility of these small proteins by use of antibody labeling.

scholarly journals Involvement of deoxyribonucleic acid polymerase β in nuclear deoxyribonucleic acid synthesis

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Dna Polymerase Beta ◽  
Cell Nuclei ◽  
High Molecular Weight Dna ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.

scholarly journals Identification of the proteins responsible for SAR DNA binding in nuclear matrix of Cucurbita pepo.

1995 ◽  
Vol 42 (2) ◽  
pp. 171-176
Author(s):  
R Rzepecki ◽  
E Markiewicz ◽  
J Szopa
Keyword(s):  
Dna Binding ◽  
Cucurbita Pepo ◽  
Standard Procedure ◽  
Cell Nuclei ◽  
Mobility Shift ◽  
Gel Mobility Shift Assay ◽  
Mobility Shift Assay ◽  
Pre Treatment ◽  
Gel Mobility Shift ◽  
Triton X

The nuclear matrices from White bush (Cucurbita pepo var. patisonina) cell nuclei have been isolated using three methods: I, standard procedure involving extraction of cell nuclei with 2 M NaCl and 1% Triton X-100; II, the same with pre-treatment of cell nuclei with 0.5 mM CuSO4 (stabilisation step); and III, method with extraction by lithium diiodosalicylate (LIS), and compared the polypeptide pattern. The isolated matrices specifically bind SAR DNA derived from human beta-interferon gene in the exogenous SAR binding assay and in the gel mobility shift assay. Using IgG against the 32 kDa endonuclease we have found in the DNA-protein blot assay that this protein is one of the proteins binding SAR DNA. We have identified three proteins with molecular mass of 65 kDa, 60 kDa and 32 kDa which are responsible for SAR DNA binding in the gel mobility shift assay experiments.

scholarly journals Autoradiographic studies of DNA and histone synthesis in successive differentiation stages of pollen grain in Hyacinthus orientalis L.

2014 ◽  
Vol 50 (3) ◽  
pp. 367-380 ◽  
Author(s):  
Elżbieta Bednarska
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Vegetative Cell ◽  
Developmental Stages ◽  
Generative Cell ◽  
Pollen Grain ◽  
Cell Nuclei ◽  
Hyacinthus Orientalis ◽  
Histone Synthesis ◽  
Generative Cells

DNA and histone synthesis in five consecutive morphological stages of <em>Hyacinthus orientalis</em> L. pollen grain differentiation were studied autoradiographically. DNA synthesis was found to occur in both the generative and the vegetative cell. DNA replication in the generative cell took place when the generative cell was still adhered to the pollen grain wall but already devoid of callose wall. DNA synthesis in the generative cell slightly preceded that in the vegetative cell. Histones were synthesized in phase S of the generative and vegetative cell. In the generative cell histone synthesis also continued at a lower level after completion of DNA replication. In the developmental stages under study the nuclei of the generative cells were decidedly richer in lysine histones than vegetative cell nuclei.

scholarly journals K+/H+-antiporter nigericin arrests DNA synthesis in Ehrlich ascites carcinoma cells

1989 ◽  
Vol 86 (17) ◽  
pp. 6626-6629 ◽  
Author(s):  
L B Margolis ◽  
Y u Novikova I ◽  
I A Rozovskaya ◽  
V P Skulachev
Keyword(s):  
Membrane Potential ◽  
Dna Synthesis ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Ehrlich Ascites Carcinoma ◽  
Carcinoma Cells ◽  
Low Concentrations ◽  
Ehrlich Ascites ◽  
Inhibition Of Dna Synthesis ◽  
Dna Synthesis Inhibition

Acidification of the cytoplasm of Ehrlich ascites carcinoma cells to pH 6.3 arrests DNA synthesis in these cells. Such an effect can be achieved by incubating the cells at pH 6.2 or by adding low concentrations of the K+/H+ antiporter, the antibiotic nigericin, at neutral pH. Glucose and anaerobiosis potentiate the nigericin effect. The inhibition of DNA synthesis by nigericin occurs without any significant decrease in the ATP concentration and in the mitochondrial membrane potential. The DNA synthesis inhibition is caused neither by a decrease in the intracellular [K+] nor by an increase in the intracellular [Na+] accompanying the nigericin effect (at least at low concentrations of the antibiotic). Nigericin should thus be regarded as a type of a cytostatic primarily affecting intracellular pH.

scholarly journals Use of PG-21 immunocytochemistry to detect androgen receptors in the songbird brain.

1996 ◽  
Vol 44 (9) ◽  
pp. 1075-1080 ◽  
Author(s):  
G T Smith ◽  
E A Brenowitz ◽  
G S Prins
Keyword(s):  
Androgen Receptor ◽  
Neural Plasticity ◽  
Behavioral Plasticity ◽  
Androgen Receptors ◽  
Brain Regions ◽  
Cell Nuclei ◽  
Low Concentrations ◽  
Song Control System ◽  
Song Control ◽  
Avian Song

The avian song control system is an excellent model in which to study the effects of gonadal steroid hormones on neural and behavioral plasticity. Several of the brain regions that control song behavior concentrate androgens and/or estrogens. Investigations of the distribution and regulation of androgen receptors have been limited by the lack of a reliable immunocytochemical method to detect androgen receptors in the songbird brain. We describe a protocol by which the PG-21 polygonal antibody to the rat androgen receptor can be used to label androgen receptor-containing cells in the songbird brain. By treating songbirds of several species with testosterone 90 min before sacrifice and by using relatively low concentrations (0.5 0.75 microg/ml) of PG-21 antibody to reduce nonspecific background staining, we were able to obtain strong specific labeling of cell nuclei in androgen-sensitive brain regions. This technique will facilitate the study of the role of androgens in mediating neural plasticity in the avian brain. Testosterone pretreatment may also facilitate the use of this antibody to label androgen receptors in tissues from a wide array of nonmammalian species.

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