The analysis of 40 kDa nuclear protein, p40, in interphase cells and mitotic cells

1993 ◽  
Vol 106 (3) ◽  
pp. 741-748 ◽  
Author(s):  
Y. Kaneda ◽  
K. Kinoshita ◽  
M. Sato ◽  
K. Tanaka ◽  
Y. Kaneda
Keyword(s):  
Monoclonal Antibody ◽  
Nuclear Envelope ◽  
Nuclear Protein ◽  
Dnase I ◽  
High Salt ◽  
The Other ◽  
Interphase Nuclei ◽  
Interphase Cells ◽  
Detergent Treatment ◽  
Sequential Treatments

We previously reported that the monoclonal antibody M108 recognized a 40 kDa protein both in the nucleus and the cytoplasm. This nuclear 40 kDa antigen was located in the nuclear envelope in interphase cells and in the perichromosomal region during mitosis. Now, we have analyzed this nuclear 40 kDa protein (p40) further, through morphological and biochemical approaches. At the beginning of mitosis, the perinuclear p40 detached from the nuclear envelope and moved to surround the condensing chromatin, while in the late stage of mitosis, the perichromosomal p40 moved back to the reassembled nuclear envelope. Most of the perichromosomal p40 on the metaphase chromosome was solubilized only by DNase I treatment, not by either high salt or detergent treatment. On the other hand, the perinuclear p40 was not solubilized by DNase1 alone, or high salt detergent alone. Sequential treatments with DNase I and high salt detergent were required to extract p40 in interphase nuclei. These results suggest that p40 was associated both with the nuclear envelope and chromatin DNA in interphase nuclei, while it bound only to chromatin DNA in mitosis.

scholarly journals PROPHASING OF INTERPHASE NUCLEI AND INDUCTION OF NUCLEAR ENVELOPES AROUND METAPHASE CHROMOSOMES IN HELA AND CHINESE HAMSTER HOMO- AND HETEROKARYONS

1974 ◽  
Vol 62 (1) ◽  
pp. 104-113 ◽  
Author(s):  
Yoshitaka Obara ◽  
Lee S. Chai ◽  
Herbert Weinfeld ◽  
Avery A. Sandberg
Keyword(s):  
Nuclear Envelope ◽  
Hela Cells ◽  
Interphase Nucleus ◽  
Chinese Hamster ◽  
Binucleate Cell ◽  
Metaphase Chromosomes ◽  
Interphase Nuclei ◽  
Multinucleate Cells ◽  
Interphase Cells ◽  
Nuclear Envelope Formation

Fusing human HeLa metaphase cells with HeLa interphase cells resulted within 30 min in either of two phenomena in the resultant binucleate cell: either prophasing of the interphase nucleus or formation of a normal-appearing nuclear envelope around the metaphase chromosomes. The frequency of either occurrence was strongly dependent on environmental pH. At pH's of 6.6–8.0, prophasing predominated; at pH 8.5 nuclear envelope formation predominated. Additionally, the frequencies of the two events in multinucleate cells depended on the metaphase/interphase ratio. When the ratio was 0.33 nuclear envelope formation predominated; when it was 2.0 prophasing predominated. In their general features, the results with fused HeLa cells resembled those reported earlier with fused Chinese hamster Don cells. However, the results provided an indication that between pH 6.6 and 8.0 the HeLa metaphase cells possessed a much greater capacity than the Don metaphase cells to induce prophasing. Fusion of Don metaphase cells with HeLa interphase cells or of Don interphase cells with HeLa metaphase cells at pH 8.0 resulted in nuclear envelope formation or prophasing in each kind of heterokaryon. As in the homokaryons, the frequencies of the two events in the heterokaryons depended on the metaphase/interphase ratio. The statistics of prophasing and nuclear envelope formation in the homo- and heterokaryon populations were consistent with the notion that disruption or formation of the nuclear envelope depends on the balance attained between disruptive and formative processes.

scholarly journals INDUCTION OF NUCLEAR ENVELOPES AROUND METAPHASE CHROMOSOMES AFTER FUSION WITH INTERPHASE CELLS

1971 ◽  
Vol 51 (1) ◽  
pp. 104-115 ◽  
Author(s):  
Tatsuro Ikeuchi ◽  
Mitsuyoshi Sanbe ◽  
Herbert Weinfeld ◽  
Avery A. Sandberg
Keyword(s):  
Nuclear Envelope ◽  
Sendai Virus ◽  
Chinese Hamster ◽  
Formation Factor ◽  
Electron Microscopic ◽  
Metaphase Chromosomes ◽  
Interphase Nuclei ◽  
Multinucleate Cells ◽  
Interphase Cells ◽  
Nuclear Envelope Formation

The process of cellular fusion induced by Sendai virus in Chinese hamster cells (Don line) afforded us the opportunity to study nuclear envelope formation around metaphase sets in the presence of interphase nuclei, when chromosome pulverization failed to occur in such multinucleate cells. Morphologically, the enveloped metaphase chromosomes resembled a normal telophase nucleus, though minor differences prompted us to call it telophase-like. Electron microscopic observations demonstrated that the membranes enveloping the chromosomes appeared to be identical with a normal nuclear envelope. The longer the cells were incubated with Colcemid before fusion, the higher was the number of cells with telophase-like nuclei and the lower the percentage of cells with pulverizations. Furthermore, the number of pulverizations bore a somewhat direct relationship to the ratio of metaphase to interphase nuclei in multinucleate cells, and the number of telophase-like nuclei was inversely proportional to this ratio. A hypothesis is advanced in which a balance between the activities of a chromosome pulverization factor and a nuclear envelope formation factor, the former in metaphase cells and the latter in interphase cells, is decisive as to the nature of morphologic events observed in virus-induced fused cells.

scholarly journals Immunorecognition of the active form of the oestrogen receptor by using a monoclonal antibody

1985 ◽  
Vol 230 (1) ◽  
pp. 203-210 ◽  
Author(s):  
N Giambiagi ◽  
J R Pasqualini
Keyword(s):  
Monoclonal Antibody ◽  
Oestrogen Receptor ◽  
Active Form ◽  
Sedimentation Coefficient ◽  
High Salt ◽  
The Other ◽  
Cytosol Fraction ◽  
Reduced Pressure ◽  
Receptor Complexes ◽  
Beta Form

In previous studies, two forms (alpha and beta) of the oestrogen receptor, with different immunological characteristics, were observed in the cytosol fraction of fetal guinea-pig uterus, by using a monoclonal antibody to the human oestrogen receptor (D547Sp gamma). Only the alpha form was recognized by the antibody, shifting its sedimentation coefficient in high-salt sucrose gradients. The present work investigated the effect of several factors (time, temperature, high salt concentrations and Na2MoO4) on the interconversion of these two forms. Only the beta form was observed when cytosol was incubated with oestradiol for only 2-3 h, but 20 h later, 40-60% of the total oestradiol-receptor complexes were found as the alpha form. The transformation from the beta to the alpha form was accelerated by temperature (25 degrees C, 15 min) and exposure to high salt concentrations (0.4 M-KCl). On the other hand, Na2MoO4 completely blocked the transformation induced by time and temperature, but had little effect on that induced by KCl. The appearance of the alpha form always correlated with an increase in receptor binding to nuclei and DNA-cellulose. Finally, it was found that the isolated beta form, recovered from the gradient, was transformed into the alpha form after overnight dialysis under reduced pressure. The present data suggest that the alpha form, which is recognized by the monoclonal antibody, is the activated form of the oestrogen receptor.

A monoclonal antibody recognizes a human nuclear protein resembling Xenopus oocyte nucleoplasmin

1987 ◽  
Vol 87 (5) ◽  
pp. 713-722
Author(s):  
J. Lord ◽  
G. Brown ◽  
C. Bunce ◽  
P. Nathan
Keyword(s):  
Monoclonal Antibody ◽  
Xenopus Oocytes ◽  
Nuclear Protein ◽  
Chinese Hamster ◽  
Nucleosome Assembly ◽  
3T3 Cells ◽  
Nucleosome Assembly Protein ◽  
Subunit Molecular Weight ◽  
Interphase Cells ◽  
A Subunit

The monoclonal antibody 10BG2 (IgG3) was derived from a mouse immunized with human pre B cells. In immunofluorescence studies the antibody revealed a human nuclear-associated determinant, which in interphase cells was entirely restricted to the nucleus. In metaphase cells 10BG2 antigen was detected throughout the cytoplasm with intensified staining at the periphery of chromosomes. 10BG2 antibody stained all human normal and transformed cells tested. In contrast, the antibody did not stain mouse 3T3 cells or Chinese hamster ovary (CHO) cells. Electrophoresis under denaturing and non-denaturing conditions revealed the 10BG2 antigen to be a 130 X 10(3) to 140 X 10(3) Mr protein with a subunit molecular weight of 29.5 X 10(3). This suggests the protein is at least a tetramer. On two-dimensional gels the 10BG2 antigen had a streaked appearance and separated into two isoelectric variants (pI5.2, 5.4). The protein was also shown to be phosphorylated and thermostable, and remained in solution at pH 3.6. 10BG2 antigen was highly soluble in aqueous buffers and co-migrated on non-denaturing gels with the nucleosome-assembly protein, nucleoplasmin, purified from Xenopus oocytes. The similarity of 10BG2 antigen to nucleoplasmin is discussed.

scholarly journals A monoclonal antibody against the nucleus reveals the presence of a common protein in the nuclear envelope, the perichromosomal region, and cytoplasmic vesicles.

1987 ◽  
Vol 104 (1) ◽  
pp. 1-7 ◽  
Author(s):  
M Wataya-Kaneda ◽  
Y Kaneda ◽  
T Sakurai ◽  
H Sugawa ◽  
T Uchida
Keyword(s):  
Monoclonal Antibody ◽  
Nuclear Envelope ◽  
Mammalian Cells ◽  
Immunoblot Analysis ◽  
Antigenic Determinants ◽  
Reactive Site ◽  
Premature Chromosome Condensation ◽  
Cytoplasmic Vesicles ◽  
Interphase Cells ◽  
Immunofluorescence Studies

A monoclonal antibody that recognizes antigenic determinants on the nucleus of cultured mammalian cells was isolated. Immunofluorescence studies using this antibody showed that the recognized antigen was present not only on the nucleus but also in cytoplasmic vesicles of interphase cells and in the perichromosomal region of mitotic cells. Premature chromosome condensation analysis showed that the reactive site for this monoclonal antibody could be detected in the perichromosomal region during the G2 and M phases, but not during the G1 and S phases. Finally, immunoblot analysis showed that this monoclonal antibody prepared against the nucleus recognized a protein of approximately 40 kD both in the cytoplasm and in the perichromosomal regions.

Telomere arrangement in differentiated interphase cells of Allium cepa: a function of chromosome arm lengths at anaphase–telophase

1983 ◽  
Vol 25 (5) ◽  
pp. 478-486 ◽  
Author(s):  
Catharine P. Fussell
Keyword(s):  
Nuclear Envelope ◽  
Allium Cepa ◽  
Interphase Nucleus ◽  
Interphase Nuclei ◽  
Meristematic Cells ◽  
Differentiated Cells ◽  
Allium Cepa L ◽  
Interphase Cells ◽  
Chromosome Arm ◽  
Latitudinal Band

The arrangement of telomeres in eight types of differentiated Allium cepa L. interphase cells was studied to find whether the distribution pattern varies with differentiation as centromere distribution appears to in differentiated cells of mice and Triturus vulgaris L. The results show that telomere positions and groupings in A. cepa are essentially the same in differentiated and meristematic interphase nuclei; telomeres, which are roughly paired, are arranged in a telophase configuration along one side of the nucleus. Thus telomeres appear to maintain the same basic arrangement in differentiated and in meristematic cells. Comparison of chromosome arm lengths and interphase telomere positions suggests that interphase telomere arrangements are a function of chromosome arm lengths at the time the nuclear envelope forms. Such an arrangement would place homologous telomeres in the same latitudinal band of the interphase nucleus.

Colocalization of a high molecular mass phosphoprotein of the nuclear matrix (p255) with spliceosomes

1995 ◽  
Vol 108 (5) ◽  
pp. 1873-1882 ◽  
Author(s):  
S. Bisotto ◽  
P. Lauriault ◽  
M. Duval ◽  
M. Vincent
Keyword(s):  
Monoclonal Antibody ◽  
Nuclear Matrix ◽  
Nuclear Protein ◽  
High Ionic Strength ◽  
High Molecular Mass ◽  
Splicing Factor ◽  
Nuclear Speckles ◽  
Ionic Detergents ◽  
Interphase Cells ◽  
Nuclear Structures

It was previously demonstrated that monoclonal antibody CC-3 binds to a phosphorylation dependent epitope present on a 255 kDa nuclear protein (p255). We show here that in interphase cells, p255 distributes to typical nuclear speckles that correspond to the localization of spliceosome components as revealed by antibodies to the m3G cap of snRNAs or to the non-snRNP splicing factor SC-35. Immunofluorescence and immunoblot studies indicated that p255 is resistant to extraction with non-ionic detergents, nucleases and high ionic strength buffers and may thus be defined biochemically as a nuclear matrix phosphoprotein. To determine the nature of the association of p255 with the nuclear structure, its distribution was studied at different stages of the cell cycle and after the cells were treated with nucleases or heat shocked. We found that the antigen diffused into the cytoplasm during metaphase but was reorganized into cytoplasmic speckles during anaphase-telophase transition, where it colocalized with SC-35. Nuclear matrix preparations that were digested with DNases and RNases showed that interphasic p255 still localized to nuclear speckles even though snRNA and snRNP antigens were removed. Heat-shocked cells labelled with monoclonal antibody CC-3 exhibited more rounded and less interconnected speckles, identical to those decorated by anti-SC-35 antibody under such conditions. These results indicate that p255 and SC-35 are present in the same nuclear structures, to which they are more tightly bound than the snRNP antigens. They further suggest that both proteins are implicated in spliceosome assembly or attachment.

Compound Symbiosis in Peridinium Balticum

1973 ◽  
Vol 31 ◽  
pp. 500-501
Author(s):  
R. N. Tomas
Keyword(s):  
Endoplasmic Reticulum ◽  
Nuclear Envelope ◽  
The Other ◽  
Exact Nature ◽  
Symbiotic Relationship

Peridinium balticum appears to be unusual among the dinoflagellates in that it possesses two DNA-containing structures as determined by histochemical techniques. Ultrastructurally, the two dissimilar nuclei are contained within different protoplasts; one of the nuclei is characteristically dinophycean in nature, while the other is characteristically eucaryotic. The chloroplasts observed within P. balticum are intrinsic to an eucaryotic photosynthetic endosymbiont and not to the dinoflagellate. These organelles are surrounded by outpocketings of endoplasmic reticulum which are continuous with the eucaryotic nuclear envelope and are characterized by thylakoids composed of three apposed lamellae. Girdle lamellae and membranebounded interlamellar pyrenoids are also present. Only the plasmalemma of the endosymbiont segregates its protoplast from that of the dinophycean cytoplasm. The exact nature of this symbiotic relationship is at present not known.

scholarly journals Possible Future Monoclonal Antibody (mAb)-Based Therapy against Arbovirus Infections

2013 ◽  
Vol 2013 ◽  
pp. 1-21 ◽  
Author(s):  
Giuseppe Sautto ◽  
Nicasio Mancini ◽  
Giacomo Gorini ◽  
Massimo Clementi ◽  
Roberto Burioni
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Side Effects ◽  
Antiviral Activity ◽  
The Other ◽  
Viral Escape ◽  
Passive Immunotherapy ◽  
Medical Need

More than 150 arboviruses belonging to different families are known to infect humans, causing endemic infections as well as epidemic outbreaks. Effective vaccines to limit the occurrence of some of these infections have been licensed, while for the others several new immunogens are under development mostly for their improvements concerning safety and effectiveness profiles. On the other hand, specific and effective antiviral drugs are not yet available, posing an urgent medical need in particular for emergency cases. Neutralizing monoclonal antibodies (mAbs) have been demonstrated to be effective in the treatment of several infectious diseases as well as in preliminaryin vitroandin vivomodels of arbovirus-related infections. Given their specific antiviral activity as well-tolerated molecules with limited side effects, mAbs could represent a new therapeutic approach for the development of an effective treatment, as well as useful tools in the study of the host-virus interplay and in the development of more effective immunogens. However, before their use as candidate therapeutics, possible hurdles (e.g., Ab-dependent enhancement of infection, occurrence of viral escape variants) must be carefully evaluated. In this review are described the main arboviruses infecting humans and candidate mAbs to be possibly used in a future passive immunotherapy.

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