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B K Vig

Publications and source records attributed to B K Vig.

At least 37 records · Page 2Linked to original sources
PubMed1993

Cytogenetic changes in primary, immortalized and malignant mammalian cells.

Some chromosomes in transformed rat cells and somatic cell hybrids fail to display the presence of kinetochore proteins as detected by antikinetochore antibodies. Such chromosomes (K- chromosomes) may constitute a novel mechanism for the genesis of aneuploidy. We have analyzed primary, immortalized and malignant mammalian cells for the presence of kinetochore proteins and micronuclei. Our results suggest a correlation of the K- chromosome and micronucleus frequency with the variability in chromosome number. Upon in situ hybridization with the minor satellite and alpha satellite sequences some K- chromosomes showed a signal. This indicates that the observed lack of kinetochores is not necessarily due to a lack of centromeric DNA. We conclude that dislocated K- chromosomes may become incorporated into micronuclei which are prone to loss. Such events would be associated with the generation of aneuploidy.

3T3 Cells↗
PubMed1993

Chromosomes lacking kinetochore proteins: their meta-anaphase location suggests potential malsegregation.

We have previously reported that a rare chromosome may not carry the kinetochore protein complex--the CENtromere Proteins or CENPs. These chromosomes should not bind to spindle microtubules and, hence, should be found peripheral to the meta-anaphase arrangement exhibited by the chromosomes which do carry CENPs. This communication shows that this actually is the case. When 3T3 mouse cells were not treated with colcemid or hypotonic, the kinetochore-lacking (K-) as well as kinetochore-bearing (K+) chromosomes were found off the spindle zone. When the spindle is disrupted with mild hypotonic treatment or by colcemid, the frequency of K- chromosomes remains unchanged. However, even mild disruption of the spindle with hypotonic treatment increases the frequency of off-lying K+ chromosomes significantly. These data indicate that K- chromosomes do not bind to the spindle and, hence, are a factor in the genesis of aneuploidy. A considerable proportion of K- chromosomes carry the putative centromere DNA indicating that these are not acentric fragments. Since the CREST serum used recognizes all essential kinetochore proteins, the K- centromeres must also lack all essential CENPs.

3T3 Cells↗
PubMed1992

Formation of primary constriction and heterochromatin in mouse does not require minor satellite DNA.

Whereas the major satellite fraction in mouse extends its domain from the centromere to the distal end of the pericentric heterochromatin, the minor satellite DNA is present specifically in the centromere or primary constriction. We hybridized the biotinylated minor satellite sequence to L929 cells of mouse origin. The sequence hybridized to all chromosomes. Whereas hybridization was detected on all active centromeres, the inactive centromeres in certain dicentrics did not show any signal. This satellite, however, was detected in all inactive centromeres in a heptacentric chromosome. The intensity of fluorescence on the inactive centromeres of the heptacentric was similar to that present on the active centromeres. Several heterochromatin blocks, which were not associated with any centromere, were also found to lack hybridization with the minor satellite. The inactive centromeres, whether carrying the minor satellite DNA fraction or not, generally do not react with the antikinetochore antibodies present in the scleroderma serum. These studies are interpreted to show that (1) the primary constriction in mouse can be formed without the participation of minor satellite, (2) heterochromatin in mouse may constitute without this fraction, (3) the major and minor satellite may not be interspersed but are joined at some defined boundary, and (4) the binding of CENP-B does not depend upon the quantity of minor satellite or the number of CENP boxes present in the inactive centromeres.

Animals↗
PubMed1991

Cytogenetic variability and kinetochore proteins. Comparison among populations derived from single-cell cultures.

This study reports comparative changes in five sub-lines obtained from a transformed culture of rat cerebral origin. Two of the lines were obtained from the original cell population while three others were raised from single-cell cultures. The comparative study was carried out on the DNA content and several cytogenetic parameters including variability in chromosome number, anaphase bridges, acentric fragments, chromosomes without detectable kinetochore proteins, and the frequency of micronuclei in these five lines. All cell lines, including the single-cell-derived clones expressed considerable variability in all aspects. One interesting aspect is the evolution of a chromosome with compound centromere, which is present only in two cell lines. The data indicate that the clone derived from a single cell does not maintain uniformity and even single cells have some sort of inherent potential to generate extreme variability. Some numerical variability may be attributed to a new phenomenon of a lack of kinetochore proteins seen on some chromosomes.

Anaphase↗
PubMed1991

Centromeres without kinetochore proteins. Another mechanism for origin of aneuploidy in neoplasia.

Centromeres of all chromosomes in normal cells exhibit kinetochore proteins detectable by antikinetochore antibodies. The present communication reports that some chromosomes in a transformed cell line of rat cerebral origin fail to deposit kinetochore proteins at their centromeres. These chromosomes may not undergo normal anaphase segregation and may be either lost or enter one or the other daughter cell. The observation that some chromosomes may be without detectable kinetochore proteins suggests a noval mechanism for origin of aneuploidy in transformed and neoplastic cells.

Aneuploidy↗
PubMed1991

Kinetochore proteins, peripheral location of chromosomes and nuclear budding: another look at the genesis of aneuploidy.

Some chromosomes in the rat do not have detectable levels of kinetochore proteins as determined by antikinetochore antibody, which may constitute a mechanism for the genesis of aneuploidy. An analysis of three mouse cell lines for the presence of kinetochore proteins showed that some chromosomes lacked these proteins, i.e. are akinetochoric by this criterion. This phenomenon may contribute to the variability of chromosome number that is characteristic of such lines. In some cells there appears to be en masse detachment of kinetochores. Several kinetochore-bearing chromosomes were also observed to be located peripherally to the spindle. These might be associated with the genesis of aneuploidy. In one cell line an apparently new phenomenon of nuclear budding was observed. In this case, one to several chromosomes appear to be pushed out of the nucleus in bud-like structures. These were surrounded by a nuclear membrane and appeared to detach from the main nucleus. Perhaps these structures eventually break off as micronuclei and, hence, would also be associated with the genesis of aneuploidy.

Aneuploidy↗
PubMed1991

Lack of detectable kinetochores on some chromosomes in mouse x human hybrids.

When treated with an anti-kinetochore antibody present in the sera of scleroderma (var. CREST) patients, most chromosomes exhibit kinetochore dots at the position of the centromere. In this paper we report that some chromosomes in the mouse x human somatic cell hybrid fail to show these dots. In the early passages in a hybrid, HYG-1, the frequency of such chromosomes was higher (0.85%) than in later passages (0.45%) studied after five months of continuous culturing. In parallel, the mean number of human chromosomes in the hybrid also dropped. The somewhat hypodiploid parental cell lines, when similarly treated, showed only a rare chromosome without kinetochore dots. Immunoblots of the proteins showed that the sera used for kinetochore detection recognized all major centromere proteins (CENPs). Electron microscopy of some offlying metaphase chromosomes in another hybrid, HR61, exhibited a lack of trilamellar kinetochores. This study suggests that akinetochoric chromosomes might provide a novel mechanism responsible for chromosome loss and genesis of aneuploidy. In early passages, some cells in the hybrid showed detached kinetochores. These autonomous kinetochores could be seen in clusters and involved some mouse chromosomes also. Potential significance of these autonomous kinetochores in generating compound centromeres is discussed.

Animals↗
PubMed1990

Centromere separation. Early replication of repetitive DNA associated with inactive centromeres.

Four types of stable dicentric and one octacentric chromosomes from mouse brain tumor cells and L-929 cells were analyzed for the timing of replication of repetitive deoxyribonucleic acid (DNA) located in the centric and pericentric regions associated with active versus inactive centromeres. The repetitive DNA present in the heterochromatin blocks of inactive centromeres replicates much earlier than similar DNA associated with the active centromeres. The former appears to replicate during early to mid S when several euchromatic segments are still replicating. There seems to be little or no overlap in the timing of replication of the repetitive DNA present in the vicinity of prematurely separating centromeres (which are accessory and nonfunctional) and those that separate at meta-anaphase junction (which are the functional centromeres). In the absence of any information about the mechanism(s) controlling initiation and completion of DNA synthesis in the two types of heterochromatic blocks, the differential timing of replication of the DNA with similar base composition remains an enigma.

Animals↗
PubMed1989

Evolution of compound centromeres. A new phenomenon.

A new type of centromere aberration in a transformed cell line of rat cerebral endothelial origin is described. These cells exhibit normal monocentric, dicentric, and multicentric chromosomes. The centromeres in dicentrics and multicentrics express variable locations along the chromosome. The centromeres in some of the multicentrics are located next to each other, with small intervening noncentromeric chromatin. In others, the centromeres appear to be in the immediate vicinity of each other with no evidence of intervening chromatin. This organization of the centromeres results in what appears to be a compound centromere composed of some multiples of single centromeres. All centromeres deposit kinetochore proteins that respond to kinetochore antibody. This evidence and that obtained from electron microscopy permits the conclusion that various centromeres/kinetochores in the compound structure are functional. The study presented here points to the existence of compound large centromeres--a novel phenomenon in cytogenetics--that may be prevalent in cancer cells. In the present cell line these regions appear as long, neck-like structures in some chromosomes and may be similar to some in vivo situations such as the X in Indian muntjac.

Animals↗
PubMed1989

Sequence of centromere separation: characterization of multicentric chromosomes in a rat cell line.

The B1 cell line of rat cerebral endothelium origin exhibits several dicentric and multicentric chromosomes. These chromosomes, unlike multicentrics in mouse (Vig and Zinkowski 1986) do not show premature centromere separation. All centromeres deposit kinetochore proteins and appear to be functional. Even the centromeres which fail to migrate to the poles during anaphase and make side arm bridges bind to spindle microtubules. Some multicentric chromosomes show kinetochores spaced apart with intervening stretches of euchromatin while others are located adjacent to each other thus exhibiting tandem repeats and forming a "compound" kinetochore (Brinkeley et al. 1984). Also, unlike mouse multicentric chromosomes in which different pericentric regions and the centromeres replicate at different times, the rat chromosomes appear to replicate all pericentric and centric regions in a given multicentric simultaneously. The present studies indicate that centromeres in rat and mouse replicate during the last part of the S-phase and in continuation with the pericentric heterochromatin.

Animals↗
PubMed1989

Sequence of centromere separation: separation in a quasi-stable mouse-human somatic cell hybrid.

A quasi-stable mouse-human hybrid cell line, HR61, containing between one and ten human chromosomes was analyzed for the sequence of centromere separation. The purpose was to determine which genome of the two initiates centromere separation first. The data clearly indicate that the separation of centromeres of the human genome is not only initiated but is completed before any centromeres from the mouse chromosomes start splitting into daughter units. The information on whether uniparental chromosome loss results from a lack of deposition of kinetochore proteins was equivocal. The human genome also completes its DNA replication before the mouse genome does. Our studies, therefore, show that the timing of centromere separation is tightly linked to the completion of replication of DNA. At least in this cell line the segregant genome is not the one which exhibits delayed DNA replication.

Animals↗
PubMed1989

Centromere structure and function in neoplasia.

The mammalian centromere plays an essential role in maintenance of diploidy in the cell. It is therefore imperative that we understand the structure and function of the mammalian centromere in order to plan strategy to control the incidence of aneuploidy and resultant malformations of the nonneoplastic as well as neoplastic tissues. Even though considerable information is available about the structure and some functional aspects of centromeres of lower eukaryotes such as yeast, the structure of the mammalian centromere is still a matter of conjecture limited to an understanding of the base composition of the alphoid sequences putatively located in the centromeric DNA of higher apes. We do, however, have a better understanding of the structure and role of the kinetochore. In all eukaryotes analyzed so far, the centromeres in a given genome separate in a sequential manner dependent upon the time of replication of pericentric and centromeric DNA. Some chromosomes, generally found in neoplastic cells, that carry more than one centromere show premature separation of the accessory centromeres. These centromeres and the associated pericentric regions replicate their DNA in an earlier part of the S phase than those that show kinetochore activity; both, however, carry DNA of the same composition. The active centromeres in these chromosomes show kinetochore protein binding as detected by antikinetochore antibody; the inactive centromeres are usually devoid of these proteins. The double minutes in neoplastic cells also lack kinetochore proteins, perhaps due to a lack of any centromere. Some dicentric and multicentric chromosomes in cancer cells and transformed cell lines do not display premature centromere separation. In these chromosomes, all centromeric sites show kinetochore proteins and all centromeric regions replicate their DNA simultaneously. These chromosomes also exhibited meiotic-like behavior of some centromeres and show postanaphase separation of some centromeres, resulting in bridges. These bridges, upon breakage and rejoining of sister chromatids, generate new multicentric chromosomes. The resulting chromosomes also exhibit formation of compound kinetochores. Some of these phenomena are novel descriptions of the centromere behavior in cancer cells. This review also discusses the role of aberrant centromere separation in human biology, providing correlates between errors of centromere separation and neoplasia.

Animals↗
PubMed1989

Sequence of centromere separation: kinetochore formation and DNA replication in dicentric chromosomes showing premature centromere separation in rat cerebral cells.

A subpopulation of rat cerebral endothelial cells, designated B1, exhibits an array of multicentric chromosomes. Because of the formation of bridges at anaphase, this cell population produced new types of multicentrics at every cell division. These chromosomes showed kinetochore proteins at every centromeric site and all centromeric regions replicated their DNA at the end of the S phase, more or less simultaneously. A new subpopulation of cells, designated B2, obtained from the original sample frozen at Wayne State University displayed several dicentrics. In contrast to B1 these chromosomes exhibit premature centromere separation as reported for mouse and human cell lines. These B2 dicentrics show only one site of kinetochore protein deposition. The timing of DNA replication around the centric region of prematurely separating centromere is also changed similar to the earlier reported premature DNA synthesis for mouse dicentrics. These observations suggest a universality of relationship between premature centromere separation, a lack of kinetochore formation, and early replication of the centric/pericentric DNA associated with these centromeres. The cause of sudden change from activity to inactivity of these chromosomes, though interesting, is not clear.

Animals↗
PubMed1989

Micronuclei, kinetochores and hypoploidy: tests with some agents.

Micronuclei were induced by treating mouse L-cells with diethylstilboestrol, colchicine and benomyl. These micronuclei were analysed for the presence of kinetochores by using antikinetochore antibody. The three chemicals induced micronuclei which differed with regards to their (i) relative distribution per cell, (ii) relative frequency for being kinetochore positive or being kinetochore negative and (iii) overall relationship between their induction and hypoploidy which in part may originate from laggards expressing as micronuclei. The data indicate that a study of micronuclei may help determine the differences between the actions of different chemicals on the genetic apparatus.

Alkylating Agents↗
PubMed1989

Centromere separation and aneuploidy: a lesson from multicentric chromosomes.

Premature centromere separation somehow nullifies the deposition of kinetochore proteins in multicentric chromosomes, is associated with early DNA replication of the centromere and the pericentric region and results in a lack of functionality of the centromere. It is conceivable that monocentric chromosomes which show premature separation, like the X chromosome as found in elderly human females (Fitzgerald et al, 1975), may have similar properties which result in a failure of centromere function and, hence, aneuploidy. This may be one of the general mechanisms by which chromosomes malsegregate.

Aneuploidy↗
PubMed1988

Heterochromatin associated with active versus inactive centromeres of mouse replicates at different times.

A subline of mouse L-cells carries a dicentric chromosome in which one centromere always separates prematurely. This centromere is not involved in the dynamics of chromosome migration and is considered inactive. By use of anti-BRdU antibody binding to BRdU-treated chromosomes it is shown that the pericentric constitutive heterochromatin associated with the prematurely separating centromere replicates earlier than its counterpart associated with the active centromere and even before several euchromatic regions in the genome. These results point to a possible mechanism by which dicentric chromosomes segregate equationally.

Animals↗