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PubMed1975

Umbra limi: a model for the study of chromosome aberrations in fishes.

Due to the lack of information available on the effect of various clastogenic agents on the chromosomes of fishes, an in vivo cytogenetics model system was developed. The central mudminnow, Umbra limi, was chosen fro this study because of its ideal karyotype consisting of 22 large meta- and submetacentric chromosomes. Various organs of the fish were investigated to determine their suitability for chromosome preparations. The tissues of the intestines, stomach, kidneys, and gills were found to be the most suitable for clastogenic studies. Phase contrast observations were made on the chromosomes of control mudminnows and mudminnows exposed to 325 R of X-radiation. The control rate of spontaneous chromosome aberrations was found to be low (about 0.03%). In contrast, fish exposed to 325 R of X-rays had aberrations in approximately 30% of the metaphases per fish examined. An apparent increase in clumping and a decrease in the mitotic index were also noted. It was concluded that the chromosomes of Umbra limi displayed typical responses to low level radiation exposure and that this fish would be an ideal cytogenetics model for this study of induced chromosome aberrations in fishes.

Animals↗
PubMed1979

Statistical methods for classification of human chromosomes.

The basic technical facts of human cytogenetics and the laboratory methods employed in chromosome research are explained in simple terms. The main variables used to describe chromosome images are defined and discussed. Three discriminant analysis models for chromosome classification are developed: one in which each chromosome is classified in isolation, a modification in which the cell, if normal, contains 2 chromosomes of each of the 23 kinds, and a final one in which the cell is the unit of analysis instead of the chromosome. Suggestions are made to reduce the calculations involved and to take into account missing chromosomes. The problem of detection and classification of aberrative chromosomes is studied, also in relation to multiple cell analysis. Finally four relevant problems are briefly discussed: selection of metaphase spreads, selection of variables, uncertain reference classification and measurement of performance.

Chromosome Aberrations↗
PubMed1975

Retinoblastoma. A model of hereditary fragile chromosomal regions.

Direct karyotyping of tumour cells from three familial and two sporadic cases of retinoblastoma revealed the existence of a Dq- marker chromosome. The hypothesis is launched that a specific region on the long arm of one of the D chromosomes is the site of a locus which is essential for the sustained differentiation of specialized retinal tissue and may be the site of other loci essential for the maturation of other embryonic tissues. Fragility of this region and its potentiality for breakage under the influence of various environmental insults could be the basic cytological event leading to the development of sporadic retinoblastoma. Mutants at these loci, including those of sustained differentiation, could be a less common operational event whereby some variants could enhance the fragility of their respective chromosomal region and thereby explain the genetic transmission of retinoblastoma in certain families. It is common for the critical functional disruption of the locus to precede the deletion which may then be considered the terminal event in the fragile region.

Child, Preschool↗
PubMed1978

Interaction of microtubules and the mechanism of chromosome movement (zipper hypothesis). 3 Theoretical analysis of energy requirements and computer simulation of chromosome movement.

A theoretical analysis of the energy requirements and a computer analysis of a special case (symmetrical pull in one plane during anaphase), of the zipper model for chromosome movement (Bajer, 1973a,b) is presented. The conclusions are general, however, and can be applied to any stage of mitosis. It is assumed that the movement is due to a series of short lateral interactions (called 'zips') between microtubules, and it is shown that a particular zip could begin if the bending energy requirement for two microtubules is met, and could terminate when a strain restriction is exceeded. The series of zips ends when the energy requirements for initiation of a particular zip are not met. In such conditions, certain predictions concerning the behaviour of microtubules in the spindle can be made (see Conclusions). It is shown that hydrolysis of ATP or GTP can yield sufficient energy to bend microtubules as specified by the model, that insignificant quantities of the triphosphate would be utilized, and that the linearity of chromosome movement predicted by the model is consistent with the linearity observed in vitro.

Adenosine Triphosphate↗
PubMed1978

The rejoining of DNA double-strand breaks and a model for the formation of chromosomal rearrangements.

The recombination type of process, which has been proposed by Resnick to explain the rejoining of radiation-induced DNA double-strand breaks, is combined with the molecular theory of radiation action to provide a description of the formation of chromosomal rearrangments. It is shown that the majority of chromosomal aberration types found at the first mitosis after radiation can be explained on the basis of one radiation-induced DNA double-strand break in the backbone of the unineme chromatid, followed by the enzymatically controlled recombinational process for the rejoining of the double-strand break. The recombinogenic process for the repair of DNA double-strand breaks relies on the close association between the broken DNA double helix and homologous DNA. The homologous nature of repeated DNA base pair sequences is used, in this model, to explain the occurrence of chromosomal exchanges between non-homologous chromosomes. The important role which repetitive DNA plays in the formation of chromosomal rearrangements and in the distribution of 'break-points' found in radiation experiments is discussed.

Base Sequence↗
PubMed1975

Non-random chromosome distribution in radial metaphases from the chinese hamster. I. Uncultured cells.

Chromosome distribution was analyzed in uncultured radial metaphase cells (corneal epithelium, testicular mitotic cells, cells in the diakinesis, and cells in metaphase II) from the Chinese hamster. The hypothesis of random distribution was rejected at the 0.001 level (chi 2/3 = 31.6).--Homologous association was observed for two pairs of chromosomes (3 and 10) in corneal epithelial cells. It was observed for all four cell types. The chromosomes associated in four groups of similarly sized and shaped chromosomes. While group membership did not appear to vary, position within the group was highly variable.--An elevenpoint model of chromosome relationships was constructed from the data.

Animals↗
PubMed2024

Capturing chromosome conformation in Crenarchaea.

While there is a considerable body of knowledge regarding the molecular and structural biology and biochemistry of archaeal information processing machineries, far less is known about the nature of the substrate for these machineries-the archaeal nucleoid. In this article, we will describe recent advances in our understanding of the three-dimensional organization of the chromosomes of model organisms in the crenarchaeal phylum.

Chromosomes, Archaeal↗
PubMed1976

Envelope-associated folded chromosomes for Escherichia coli: variations under different physiological conditions.

The folded chromosome of Escherichia coli has been investigated under various lysis and physiological conditions. A new gradient system was devised that allows excellent separation between unlysed cells and envelope-associated and envelope-free chromosomes. Isotope incorporation experiments showed that the fraction often called "membrane-bound nucleoids" contains cell wall in addition to nucleic acids, membranes, and proteins. The amount of lysozyme added and the lysozyme digestion time were found to be important when comparing the rate of sedimentation of envelope-associated chromosomes obtained under various physiological conditions. Amino acid-starved cells were found to be much harder to lyse with lysozyme than exponentially grown cells, The difference in sedimentation coefficient of envelope-associated chromosomes described earlier (Ryder and Smith, 1974) was not detected when the latter two types of cells had been given equivalent, but not identical, lysozyme treatment such that detergent-mediated lysis proceeded at the same rate. Analysis of pulse- and uniformly labeled chromosomes from amino acid-starved cultures revealed no preferential labeling of either envelope-associated or -released nucleoids. Nor was there a difference in sedimentation coefficient between uniform and pulse-labeled envelope-associated nucleoids. These results are in disagreement with the models for chromosome replication of Worcel and Burgi (1974) and Ryder and Smith (1974), respectively. Growing cells on carbon sources poorer than glucose demonstrated that the replicating chromosomes sediment faster than the bulk of envelope-associated nucleoids. The slower the growth rate, the greater this difference became. An alternative hypothesis regarding chromosome replication and its association with the cell envelope is presented.

Amino Acids↗
PubMedDate not supplied

[Structure of chromosomal deoxyribonucleoproteins. VII. Free dna in preparation of fragmented chromatin].

Chromatin which was hydrodynamically sheared in a low ionic strength buffer lacking divalent cations (mu = 0.005) contains a heterogeneous set of DNP particles but no molecules of free DNA. The main finding is that a transference of sheared chromatin to 1-2 mM MgCl2 or to 0.1-0.2M NaCl results in the appearance of completely free DNA molecules. A salt-induced rearrangementof DNA-bound histones, but not a partial loss of them is responsible for the observed phenomenon. Formation of free DNA molecules is accompanied by aggregation of the majority of remaining DNP particles. Percentage of free DNA molecules in the chromatin which was sheared to an average DNA length of approx. 400 base pairs is increased from zero in the initial DNP sample to 8-9% in 1 mM MgCl2 and further to 30-31% of the total DNA in 0.30 M NaCl, 2 mM MgCl2. Free DNA molecules in the sheared chromatin are observed not only upon isopycnic banding of formaldehyde-fixed DNP in CsCl gradients but also in non-ionic Metrizamide gradients with either fixed or unfixed DNP samples. Process of free DNA formation is a reversible one; its direction and the equilibrium state depend in particular on the ionic conditions of the medium. Percentage of free DNA molecules in the sheared chromatin at a given ionic strength of solution is strongly decreased upon an increase of the average length of DNA in the DNP particles. Several lines of evidence suggest that free DNA molecules are formed in the sheared chromatin as a result of cooperative rearrangements of histones in salt-induced DNP aggregates. A dynamical model of chromosomal fiber is proposed on the basis of the present and earlier experimental data [1]. According to the model histones are arranged on DNA in clusters separated by stretches of free DNA. A salt-induced migration of histones along or between DNP fibers can result in unification of different clusters, thereby generating longer stretches of free DNA, the total amount of free DNA being approximately constant. Possible in vivo significance of such a dynamical structure is discussed.

Animals↗
PubMed1976

The effects of deoxyribonucleic acid secondary structure on tertiary structure.

The secondary structure of supercoiled DNA was varied by changes in ionic strength. For I = 0.075-0.4 the structure remained in the previously established branched form with only minor alterations in molecular dimensions. In 4M-NaCl, which induces linear DNA to change its secondary structure to the C structure and brings about an increase in the superhelix density of the molecule, no extra branches were observed on the molecules. The limiting factors that dictate supercoil structure seem to be the number and position of potential branch points and the proximity with which the two intertwining DNA strands can approach each other on the arms of the branches. This value is close to 10nm under the conditions described, and is 14-15nm at I = 0.2. It is suggested that such values should be borne in mind when models of chromosome structure are being constructed.

Bacteriophages↗
PubMed1978

dnaT, dominant conditional-lethal mutation affecting DNA replication in Escherichia coli.

Normally, bacteria cease DNA replication in the absence of protein synthesis. A variety of treatments, such as thymine starvation or a shift-up to rich medium, lead to continued DNA replication in the absence of protein synthesis. Mutants are described which always terminate replication under these conditions. These conditional lethal mutants, dnaT1 and dnaT2, contransduce with serB and dnaC. The mutation also affects cell division. All aspects of the mutant phenotype (obligatory termination of replication, temperature sensitivity of DNA replication and growth, and aberrant cell division at permissive growth temperatures) were transdominant to the wild-type phenotype. Episomes carrying the dnaT mutation appeared to be unstable. The existence of such a dominant mutation was predicted by a model of chromosome termination proposed by Kogoma and Lark (J. Mol. Biol. 94:243-256, 1975).

Bacterial Proteins↗
PubMed1977

[Genome structure in higher plants. Reassociation kinetics of DNA from Vicia faba and Vicia sativa].

The work been concerned with a study of the kinetics of reassociation of total DNA and that of the fraction of unique sequences in plants from the Vicia family, i. e. Vicia faba and Vicia sativa. The size of the genome was determined by the kinetics of reassociation of the DNA of the fraction of unique sequences and the amount of DNA per nucleus was determined cytophotometrically. It has been shown that the size of the genome expressed in C(0)t units and the size expressed in gramms are not the same which testifies to the absence of true unique genes in the genome of the species studied. The analysis of the possible methodical errors was carried out. On the basis of the data obtained a suggestion was made of a model of chromosomes organization including 12 units of polynemization for Vicia faba and 4 units for Vicia sativa.

Base Sequence↗
PubMed1975

X-ray and UV-induced chromatid aberrations: evidence for polynemic chromosomes?

Ikushima and Wolff have recently interpreted both their observation of chromatid aberrations in second and third mitoses following X-irradiation and the production of chromatid type chromosomal aberrations by UV light administered during the G1 phase of the cell cycle in terms of a polyneme model of eukaryote chromosome structure. They were led to do so, however, largely because of their X-ray data, which the interpreted as evidence for the induction of sub-chromosomal lesions (by G1 irradiation; sub-chromatid for G2 irradiation) which appear as chromatid type aberrations only in later divisions. We here report data from similar X-ray experiments in which synchronized Chinese hamster tissue culture cells were irradiated in either G1 or G2 and then scored for chromatid aberrations in their first, second and third post-irradiation mitoses. Our results do not show the effect reported by Ikushima and Wolff. We conclude that all of the data available of aberration production is compatible with a simple mononeme model of eukaryote chromosome structure.

Animals↗