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At least 1,765 records · Page 98Linked to original sources
PubMed1997

Micrococcal nuclease (endonuclease) digestion causes apoptosis and mitotic catastrophe with interphase chromosome condensation in human Chang liver cells.

Endonuclease activation causing genomic degradation is a pervasive hallmark of apoptosis and a suggested precipitating or commitment step in the suicidal process. Directly applied endonuclease activity has produced apoptotic-like effects in isolated nuclei, but not yet shown as an initiating apoptogen in whole cells. Mechanistically genomic damage inflicted by a variety of DNA-damaging agents is also known to produce mitotic catastrophe condensations characterizing cell cycle derangement. Morphological and molecular similarities between apoptosis and mitotic catastrophe have been noted, but their conjoint expressions from directly applied endonuclease activity has also not been shown. We show here micrococcal nuclease (MNase) initiating apoptosis in human Chang liver cells which expressed both apoptotic and mitotic catastrophe condensations. Genomic profiling showed (a) the two stage apoptotic sequence of large (50 kb) and small (200 bp) fragment cleavage demonstrated by pulse field and normal gel electrophoresis, respectively; (b) the sub-G1 ;apoptotic peak' with shrunken cells from flow cytometric evaluation of PI-DNA binding and laser forward scattering, (c) 3' OH termini typical of apoptotic DNA fragments labelled by terminal deoxynucleotidyl transferase (TdT)-mediated fluorescence tagging especially in the shrunken cells, and (d) positive comet assay of the apoptotic genome. Nuclear shrinkage evaluated by confocal image analysis was consistent with the apoptotic response, as was Zn2+ ion sensitivity, an established inhibitor of apoptotic expression. Endonuclease activity per se is apoptogenetic and mechanistically convergent with the mitotic catastrophe pathway in the proliferative cycle.

Journal Article↗
PubMed2003

Genetic instability in superficial bladder cancer and adjacent mucosa: an interphase cytogenetic study.

A systematic analysis of both tumors and the surrounding urothelium to help identify what lies behind the mechanism of multifocal tumor development has not yet been performed. In this study we investigated chromosome 1, 7, 9, and 17 aneusomy in 25 superficial papillary carcinomas and in 51 tissue samples taken from sites of macroscopically uninvolved urothelium surrounding the tumors, using the fluorescence in situ hybridization method. Our data demonstrated a close genetic relationship between all examined tumors and normal-appearing mucosa. Numeric aberrations of chromosomes 1, 7, 9, and 17 were found to exhibit similar patterns in all analyzed specimens, although with different frequencies.

Carcinoma, Papillary↗
PubMed2004

Functional evidence for active site location of tetrameric thymidylate synthase X at the interphase of three monomers.

Little is known about the catalytic mechanism of the recently discovered ThyX family of flavin-dependent thymidylate synthases that are required for thymidylate (deoxythymidine 5'-monophosphate) synthesis in a large number of microbial species. Using a combination of site-directed mutagenesis and biochemical measurements, we have identified several residues of the Helicobacter pylori ThyX protein with crucial roles in ThyX catalysis. By providing functional evidence that the active site(s) of homotetrameric ThyX proteins is formed by three different subunits, our findings suggest that ThyX proteins have evolved through multimerization of inactive monomers. Moreover, because the active-site configurations of ThyX proteins, present in many human pathogenic bacteria, and of human thymidylate synthase ThyA are different, our results will aid in the identification of compounds specifically inhibiting microbial growth.

Amino Acid Sequence↗
PubMed1980

Geometrical model for the energy of semicoherent interphase interfaces.

The basis for the considerations given in this paper is the O-lattice description of crystalline interfaces of Bollmann. In the development of his approach presented here, all possible interfacial planes between two crystal phases having a defined orientation relationship are considered. The energies of these interfaces are then computed in terms of the energies of the primary intrinsic dislocations. A number of modeling interactions are incorporated into this approach, and a better agreement with experimental data is thus obtained.

Journal Article↗
PubMed1987

Three-dimensional organization of Drosophila melanogaster interphase nuclei. II. Chromosome spatial organization and gene regulation.

In the preceding article we compared the general organization of polytene chromosomes in four different Drosophila melanogaster cell types. Here we describe experiments aimed at testing for a potential role of three-dimensional chromosome folding and positioning in modulating gene expression and examining specific chromosome interactions with different nuclear structures. By charting the configurations of salivary gland chromosomes as the cells undergo functional changes, it is shown that loci are not repositioned within the nucleus when the pattern of transcription changes. Heterologous loci show no evidence of specific physical interactions with one another in any of the cell types. However, a specific subset of chromosomal loci is attached to the nuclear envelope, and this subset is extremely similar in at least two tissues. In contrast, no specific interactions between any locus and the nucleolus are found, but the base of the X chromosome, containing the nucleolar organizer, is closely linked to this organelle. These results are used to evaluate models of gene regulation that involve the specific intranuclear positioning of gene sequences. Finally, data are presented on an unusual class of nuclear envelope structures, filled with large, electron-dense particles, that are usually associated with chromosomes.

Animals↗
PubMed1994

Visualization of G1 chromosomes: a folded, twisted, supercoiled chromonema model of interphase chromatid structure.

We have used light microscopy and serial thin-section electron microscopy to visualize intermediates of chromosome decondensation during G1 progression in synchronized CHO cells. In early G1, tightly coiled 100-130-nm "chromonema" fibers are visualized within partially decondensed chromatin masses. Progression from early to middle G1 is accompanied by a progressive uncoiling and straightening of these chromonema fibers. Further decondensation in later G1 and early S phase results in predominantly 60-80-nm chromonema fibers that can be traced up to 2-3 microns in length as discrete fibers. Abrupt transitions in diameter from 100-130 to 60-80 nm along individual fibers are suggestive of coiling of the 60-80-nm chromonema fibers to form the thicker 100-130-nm chromonema fiber. Local unfolding of these chromonema fibers, corresponding to DNA regions tens to hundreds of kilobases in length, reveal more loosely folded and extended 30-nm chromatin fibers. Kinks and supercoils appear as prominent features at all observed levels of folding. These results are inconsistent with prevailing models of chromosome structure and, instead, suggest a folded chromonema model of chromosome structure.

Animals↗
PubMed1965

Inducibility of thymidine kinase by thymidine as a function of interphase stage.

Thymidine may act as an inducer of thymidine kinase activity in cells of higher plants. The general response is demonstrable in a randomly developing cell population such as is found in germinating wheat embryos. If a synchronously developing cell population is studied, however, potentially inducible cells are found to be susceptible to the inductive effect of thymidine only during about 10 per cent of the G1 period, and close to the interval when thymidine kinase activity normally appears.

Cell Division↗
PubMed1967

DNA-protein binding in interphase chromosomes.

The metachromatic dye, azure B, was analyzed by microspectrophotometry when bound to DNA fibers and DNA in nuclei with condensed and dispersed chromatin. The interaction of DNA and protein was inferred from the amount of metachromasy (increased beta/alpha-peak) of azure B that resulted after specific removal of various protein fractions. Dye bound to DNA-histone fibers and frog liver nuclei fixed by freeze-methanol substitution shows orthochromatic, blue-green staining under specific staining conditions, while metachromasy (blue or purple color) results from staining DNA fibers without histone or tissue nuclei after protein removal. The dispersed chromatin of hepatocytes was compared to the condensed chromatin of erythrocytes to see whether there were differences in DNA-protein binding in "active" and "inactive" nuclei. Extraction of histones with 0.02 N HCl, acidified alcohol, perchloric acid, and trypsin digestion all resulted in increased dye binding. The amount of metachromasy varied, however; removal of "lysine-rich" histone (extractable with 0.02 N HCl) caused a blue color, and a purplish-red color (micro-peak absorption) resulted from prolonged trypsin digestion. In all cases, the condensed and the dispersed chromatin behaved in the same way, indicating the similarity of protein bound to DNA in condensed and dispersed chromatin. The results appear to indicate that "lysine-rich" histone is bound to adjacent anionic sites of a DNA molecule and that nonhistone protein is located between adjacent DNA molecules in both condensed and dispersed chromatin.

Animals↗