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At least 73 records · Page 4Linked to original sources
PubMed1957

Mononucleotides of the cell nucleus.

1. It has been demonstrated by ion exchange chromatography that the cell nucleus contains mononucleotides of adenine, guanine, cytosine, uracil, together with diphosphopyridine nucleotide, and several uridine diphosphate derivatives; the adenine nucleotides predominating in amount. Nucleotide components in the cell nucleus are in close agreement both quantitatively and qualitatively with those found in the cytoplasm. 2. In calf thymus sucrose nuclei, nucleotide monophosphates can be phosphorylated to the energy-rich triphosphate form without participation of cytoplasmic components. As to the nature of the phosphorylation, it has been shown that there exist certain differences as well as resemblances between nuclei and mitochondria. A distinctive feature of nuclear phosphorylation is that only intranuclear monophosphates seem to be phosphorylated. The process is completely inhibited by cyanide, azide, and dinitrophenol. However, certain reagents which block oxidative phosphorylation of mitochondria, namely dicumarol, Janus green B, methylene blue, and calcium ions, have no effect on phosphorylation within the nucleus. 3. The bulk of mononucleotides is preserved within thymus nuclei after their isolation in sucrose. Nucleotides are surprisingly well retained by nuclei in a sucrose medium whether or not electrolytes are present and in buffers ranging from pH 3 to 10; under all conditions sucrose is required for retention. 4. Dilute acetate in sucrose releases nucleotides from the nucleus below pH 5.1. As to the effective pH of acetate, there is a sharp boundary between pH 5.1 and pH 5.9. At pH 5.9, and above, acetate does not remove nucleotides from the nucleus. The effects of propionate, formate, and monochloroacetate on the nuclei are the same as that of acetate. 5. When nuclei are exposed to a wide variety of conditions a close correlation is found between the retention in the nucleus of nucleotides and of potassium. This suggests that both substances are part of a common complex in the cell nucleus. 6. It has been shown that upon removal of nucleotides and potassium from calf thymus sucrose nuclei by acetate, the ability to incorporate C(14)-alanine into nuclear protein is greatly impaired.

Animals↗
PubMed2001

Kinetics and mechanism of DNA uptake into the cell nucleus.

Gene transfer to eukaryotic cells requires the uptake of exogenous DNA into the cell nucleus. Except during mitosis, molecular access to the nuclear interior is limited to passage through the nuclear pores. Here we demonstrate the nuclear uptake of extended linear DNA molecules by a combination of fluorescence microscopy and single-molecule manipulation techniques, using the latter to follow uptake kinetics of individual molecules in real time. The assays were carried out on nuclei reconstituted in vitro from extracts of Xenopus eggs, which provide both a complete complement of biochemical factors involved in nuclear protein import, and unobstructed access to the nuclear pores. We find that uptake of DNA is independent of ATP or GTP hydrolysis, but is blocked by wheat germ agglutinin. The kinetics are much slower than would be expected from hydrodynamic considerations. A fit of the data to a simple model suggests femto-Newton forces and a large friction relevant to the uptake process.

Adenosine Triphosphate↗
PubMed1996

Absorbed fraction to the cell nucleus for low energy electrons.

The absorbed fraction phi to the cell nucleus for low energy electrons (0.5 keV-50 keV) was evaluated. Distributions of radioactivity within either the nucleus (nu) and the cytoplasm (cy), or on the cell membrane (mem) were considered. phi was computed as a function of the cell sizes and of the electron energy E. For a strictly intranuclear distribution, phi(nu) is close to 1 for very low energy values (E < 4 keV), i.e. the energy is totally absorbed in the cell nucleus itself. The absorbed fraction decreases when the energy increases and phi(nu) becomes less than 0.1 for E > or = 40 keV. For a cell membrane distribution, the absorbed fraction remains always less than 0.2. For very low energy electrons (E < 6 keV), phi(mem) = 0, due to the fact that the electron falls short to the nucleus target. The absorbed fraction is maximum for E ranging from 12 keV to 20 keV. For higher values of E, phi(mem) decreases when E increases. When considering a cytoplasmic distribution, the maximum absorbed fraction phi(cy) is obtained for E values ranging from 10 keV to 25 keV (phi(cy max) = 0.27). Dosimetric computations at the cellular level show that the absorbed fraction to the cell nucleus may have values ranging from 0 to 1, depending on the dimensions of the cell, the energy of the emitted electron and on the intracellular localization of the Auger emitter.

Absorption↗
PubMed1976

Synaptic events and discharge patterns of cochlear nucleus cells. II. Frequency-modulated tones.

Responses of 99 cochlear nucleus cells and 24 cochlear nerve fibers were studied with FM signals; 14 cochlear nerve fibers and 57 cochlear nucleus cells were studied at four rates of modulation and several signal intensities. Classification of FM response patterns as symmetrical, asymmetrical, or unidirectional was based on the calculation of a symmetry factor (S), which compared the number of discharges evoked by the ascending and by the descending phases of the FM sweep. Certain FM response patterns could not adequately be described by the symmetry factor along and variables of modulation rate and signal intensity had significant influence. A correspondence was found between the four response classes evoked by a steady-frequency tone burst (primarylike, buildup, onset, and pause) and the FM response pattern. Cochlear nerve fibers showed symmetrical response patterns to FM stimulation. Primarylike units were similar to eighth nerve fibers and generally showed symmetrical FM responses. Occasional eighth nerve fibers and primarylike cells developed asymmetry at the fastest rate of modulation (50 sps). Buildup units showed a variety of response patterns to FM signals. Onset units generally showed asymmetrical response patterns with the greater response occurring to the ascending than to the descending phase of the FM sweep. Pause units showed a characteristic inhibition of activity at 5 sps (rate-dependent inhibition). Of the 57 cochlear nuclear cells studied in response to FM signals, 16 were symmetrical, another 16 were symmetrical except at the fastest modulation rate, 12 were asymmetrical, 3 were unidirectional, and 10 showed complex responses to certain signal rates or intensities. It is clear the the cat cochlear with its complex cytoarchitecture is involved in the recoding of acoustic information. Some units in cochlear nucleus demonstrate differential responses to the direction and to the rate of frequency movement. Other cochlear nucleus cells respond as eighth nerve fibers and may serve as simple "relays" in transmitting information from the cochlea to higher auditory centers.

Acoustic Stimulation↗
PubMed2004

Poly(A)+ RNAs roam the cell nucleus and pass through speckle domains in transcriptionally active and inactive cells.

Many of the protein factors that play a role in nuclear export of mRNAs have been identified, but still little is known about how mRNAs are transported through the cell nucleus and which nuclear compartments are involved in mRNA transport. Using fluorescent 2'O-methyl oligoribonucleotide probes, we investigated the mobility of poly(A)+ RNA in the nucleoplasm and in nuclear speckles of U2OS cells. Quantitative analysis of diffusion using photobleaching techniques revealed that the majority of poly(A)+ RNA move throughout the nucleus, including in and out of speckles (also called SC-35 domains), which are enriched for splicing factors. Interestingly, in the presence of the transcription inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole, the association of poly(A)+ RNA with speckles remained dynamic. Our results show that RNA movement is energy dependent and that the proportion of nuclear poly(A)+ RNA that resides in speckles is a dynamic population that transiently interacts with speckles independent of the transcriptional status of the cell. Rather than the poly(A)+ RNA within speckles serving a stable structural role, our findings support the suggestion of a more active role of these regions in nuclear RNA metabolism and/or transport.

Active Transport, Cell Nucleus↗
PubMed1994

The influence of tracer localization on the electron dose rate delivered to the cell nucleus.

UNLABELLED: The radiation dose rate delivered by electron emissions of 99mTc, 123I, 111In, 67Ga and 201Tl was evaluated at the subcellular level. METHODS: Spherical models of sources were used to simulate various cellular localizations of radionuclides. These models were applied to large lymphocytes, assuming uniform distributions of radioactivity throughout the nucleus, the cytoplasm or the cell membrane surface. RESULTS: The graphs of the absorbed dose rate plotted according to the distance from the center of the cell show that the dose rate strongly depends on the subcellular distribution of the radioisotope. The absorbed dose rate D(0) at the center of the cell delivered by a constant cellular radioactivity of 99mTc, 123I, 111In, 67Ga and 201Tl is respectively 94, 21, 18, 74 and 76 times higher if the radioactivity is localized within the cell nucleus than if it is situated only on the cell membrane. D(0) for subcellular localizations was compared to D(0) obtained by assuming uniform distribution of radioactivity throughout the cell. This latter assumption may underestimate the dose rate from 2.8- to 3.2-fold if the tracer is exclusively localized within the nucleus or overestimate from 4.3- to 30-fold if the tracer is localized within the cytoplasm or on the cell membrane, depending on the radionuclide. CONCLUSION: Such findings show that the localization of radiopharmaceuticals at the subcellular level plays a crucial role in determining the actual dose delivered to the cell nucleus in diagnostic nuclear medicine procedures.

Cell Membrane↗
PubMed1987

The role of the host cell nucleus in vaccinia virus morphogenesis.

Despite the fact that cells infected with wild type vaccinia virus synthesize viral DNA and assemble progeny virus particles within the cytoplasm, the host cell nucleus is required for a productive infection. Recent evidence suggests that vaccinia virus selectively recruits components from the host cell nucleus into the cytoplasm for use by the developing virus. One of these components is the largest subunit of the cellular RNA polymerase II (Pol II).

Animals↗
PubMed1995

The target cell nucleus is not required for cell-mediated granzyme- or Fas-based cytotoxicity.

The requirement for target cell nuclei in the two apoptotic death pathways used by cytotoxic lymphocytes was tested using model effector systems in which the granzyme and Fas pathways of target damage are isolated. Mast cell tumors expressing granzymes A and B in addition to cytolysin/perforin lysed tumor target cells about 10-fold more efficiently than comparable effector cells without granzymes. Enucleated cytoplast targets derived from these cells were also lysed with a similar 10-fold effect of granzymes. In contrast to cytoplasts, effector granzyme expression did not influence lysis of red cell targets. The Fas pathway was assessed using the selected cytotoxic T lymphocyte hybridoma subline d11S, which lysed target cells expressing Fas but not those lacking Fas. Similarly, cytoplasts derived from Fas+ but not Fas- cells were also readily lysed by these effector cells. Thus, neither the nucleus itself nor the characteristic apoptotic nuclear damage associated with the two major cell death pathways used by cytotoxic lymphocytes are required for cell death per se.

Antigens, Surface↗
PubMed1999

Toward a cancer therapy with boron-rich oligomeric phosphate diesters that target the cell nucleus.

The viability of boron neutron capture therapy depends on the development of tumor-targeting agents that contain large numbers of boron-10 (10B) atoms and are readily taken up by cells. Here we report on the selective uptake of homogeneous fluorescein-labeled nido-carboranyl oligomeric phosphate diesters (nido-OPDs) by the cell nucleus and their long-term retention after their delivery into the cytoplasm of TC7 cells by microinjection. All nido-OPDs accumulated in the cell nucleus within 2 h after microinjection. However, nido-OPDs in which the carborane cage was located on a side chain attached to the oligomeric backbone were redistributed between both the cytoplasm and nucleus after 24 h of incubation, whereas nido-OPDs in which the carborane cage was located along the oligomeric backbone remained primarily in the nucleus. Furthermore, cell-free incubation of digitonin-permeabilized TC7 cells with the nido-OPDs resulted in nuclear accumulation of the compounds, thus corroborating the microinjection studies. Our observation of fluorescence primarily located in the cell nucleus indicates that nuclear-specific uptake of sufficient amounts of 10B for effective boron neutron capture therapy ( approximately 10(8)-10(9) 10B atoms/tumor cell) via nido-OPDs is achievable.

Biological Transport↗
PubMed1991

The nuclear matrix: a heuristic model for investigating genomic organization and function in the cell nucleus.

Despite significant advances in deciphering the molecular events underlying genomic function, our understanding of these integrated processes inside the functioning cell nucleus has, until recently, met with only very limited success. A major conundrum has been the "layers of complexity" characteristic of all cell structure and function. To understand how the cell nucleus functions, we must also understand how the cell nucleus is put together and functions as a whole. The value of this neo-holistic approach is demonstrated by the enormous progress made in recent years in identifying a wide variety of nuclear functions associated with the nuclear matrix. In this article we summarize basic properties of in situ nuclear structure, isolated nuclear matrix systems, nuclear matrix-associated functions, and DNA replication in particular. Emphasis is placed on identifying current problems and directions of research in this field and illustrating the intrinsic heuristic value of this global approach to genomic organization and function.

Amino Acid Sequence↗
PubMed1983

Requirement of cell nucleus for Sindbis virus replication in cultured Aedes albopictus cells.

The ability of Sindbis virus to grow in enucleated BHK-21 (vertebrate) and Aedes albopictus (invertebrate) cells was tested to determine the dependence of this virus upon nuclear function in these two phylogenetically unrelated hosts. Although both cell types could be demonstrated to produce viable cytoplasts (enucleated cells) which produced virus-specific antigen subsequent to infection. BHK cytoplasts produced a significant number of progeny virions, whereas mosquito cytoplasts did not. The production of vesicular stomatitis virus in mosquito cells was not significantly reduced by enucleation. That such a host function was not essential for vesicular stomatitis virus growth in insect cells is supported by the observation that the production of this virus by mosquito cells is not actinomycin D sensitive. This result agrees with a previously published report in which it was shown that Sindbis virus maturation in invertebrate cells is inhibited by actinomycin D, indicating a possible requirement for host cell nuclear function (Scheefers-Borchel et al., Virology, 110:292-301, 1981).

Aedes↗
PubMed2006

The road much traveled: trafficking in the cell nucleus.

Trafficking of RNA molecules and proteins within the cell nucleus is central to genome function. Recent work has revealed the nature of RNA and protein motion within the nucleus and across the nuclear membrane. These studies have given insight into how molecules find their destinations within the nucleus and have uncovered some of the structural properties of the nuclear microenvironment. Control of RNA and protein trafficking is now emerging as a physiological regulatory mechanism in gene expression and nuclear function.

Active Transport, Cell Nucleus↗
PubMed1979

Vaccinia virus replication. I. Requirement for the host-cell nucleus.

Using cytochalasin B-induced enucleation techniques, we examined the ability of vaccinia virus to replicate in the absence of the host-cell nucleus in several mammalian cell lines. It was found that virus-infected enucleated cells (cytoplasts) prepared from BSC-40, CVC, and L cells were incapable of producing infectious progeny virus. The nature of this apparent nuclear involvement was studied in detail in BSC-40 cells. Modulations designed to maximize cytoplast integrity and longevity, such as reduction of the growth temperature and initial multiplicity of infection, did not improve virus growth in cytoplasts. Sodium dodecyl sulfate-polyacrylamide gel analysis of the [(35)S]methionine pulse-labeled proteins synthesized in vaccinia virus-infected cytoplasts demonstrated that both early and late viral gene products were being expressed at high levels and with the proper temporal sequence. Vaccinia virus cytoplasmic DNA synthesis, as measured by [(3)H]thymidine incorporation, peaked at 3 h postinfection and was 70 to 90% of control levels in cytoplasts. However, in the cytoplasts this DNA was not converted to a DNase-resistant form late in infection, which was consistent with the failure to isolate physical particles from infected cytoplasts. Treatment of vaccinia virus-infected cells with 100 mug of rifampin/ml from 0 to 8 h to increase the pools of viral precursors, followed by subsequent removal of the drug, resulted in a threefold increase virus yield. This treatment had no effect on virus-infected cytoplasts. Finally, vaccinia virus morphogenesis was studied under an electron microscope in thin sections of virus-infected cells and cytoplasts which had been prepared at various times during a single-step growth cycle. It was apparent that, although early virus morphogenetic forms appeared, there was no subsequent DNA condensation or particle maturation in the cytoplasts. These results suggest that vaccinia virus requires some factor or function from the host-cell nucleus in order to mature properly and produce infectious progeny virus.

Animals↗
PubMed1997

Apoptosis induced by methylazoxymethanol in developing rat cerebellum: organization of the cell nucleus and its relationship to DNA and rRNA degradation.

We present a cytological and biochemical study of the cell death of granule cell precursors in developing rat cerebellum following treatment with the cytotoxic agent methylazoxymethanol (MAM) during the first postnatal week. The density of apoptotic figures per square millimeter progressively increases after 6, 12, 24 and 44 h of treatment, whereas cells immunoreactive for proliferating cell nuclear antigen tend to disappear in the external granular layer (EGL). DNA migration on gel electrophoresis reveals a typical ladder pattern of internucleosomal cleavage following MAM treatment, whereas gel electrophoresis of rRNA shows a conspicuous degradation of both 28S and 18S rRNAs. Ultrastructural analysis has revealed the alterations of structures containing chromatin and ribonucleoprotein (RNP) in dying cells of the EGL. The typical granular beaded configuration of the condensed chromatin changes to a denser, more homogeneous texture suggesting nucleosomal disruption. The reorganization of RNP nuclear domains is reflected by the appearance of dispersed nucleoplasmic RNP particles and the formation of a coiled-body-like structure. However, typical nuclear domains involved in the splicing of RNAs, namely interchromatin granule clusters and typical "coiled bodies", are not found in apoptotic cells. Intranuclear bundles of filaments have also been detected. In the cytoplasm, the presence of dispersed single ribosomes is an initial sign of apoptosis. The massive dispersion and disruption of ribosomes detected after 24 h and 44 h of MAM treatment is reflected by the degradation of both 28S and 18s rRNAs. These results show that MAM treatment provides a useful experimental model for the study of apoptosis in the developing central nervous system. The organization of the cell nucleus in cells undergoing apoptosis clearly reflects a disruption of the nuclear compartments involved in transcription and the processing and transport of RNA and is related to the patterns of DNA and rRNA degradation.

Animals↗
PubMed1980

Methods in ultrastructural cytochemistry of the cell nucleus.

The electron microscopical study of the cell nucleus as observed in thin sections requires the use of cytochemical methods because of the intricate pattern of the nuclear components. The in situ techniques based on electron staining and enzymatic digestion are reviewed, excluding autoradiography, cytoenzymology and immunocytochemistry. A tentative classification has been adopted according to the chemical nature of the revealed component. Thus, the staining procedures for the nucleoproteins in general, for both nucleic acids, for the proteins, and finally for the deoxyribonucleoproteins and DNA are considered separately. 1--Stains for the nucleoproteins include simple reagents such as the uranyl and lead salts which are largely used in electron microscopy but are of limited specificity. 2--A variety of methods, some of them specific, is available for the simultaneous visualization of DNA and RNA which is based on common properties: basophilia, ability to bind diaminoacridines, presence of hydroxyl groups. However, due to the recent development of specific and preferential methods for each nucleic acid, we feel that among the older methods, only rapid and simple procedures for the detection of both nucleic acids remain of interest. 3--Proteins being ubiquitous, the useful techniques must reveal subsets within the total nuclear proteins. Apart from some endogeneous enzymes, basic proteins -- practically histones -- so far represent the only group for the detection of which reliable methods exist. 4--Several techniques developed recently are available for the specific detection of DNA. In favourable cases, methods derived from the Feulgen reaction allow its visualization at a molecular level. In addition, standard procedures for the preparation of mammalian cells and tissues are described. Each staining method is at least briefly discussed, but emphasis has been placed on a small number of techniques described in detail. They comprise the EDTA regressive stain for the ribonucleoproteins, several reactions of the basic proteins and the Feulgen-like osmium ammine reaction for DNA.

Cell Nucleus↗