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Biomedical subjects

D M Prescott

Publications and source records attributed to D M Prescott.

At least 91 records · Page 5Linked to original sources
PubMed1980

Gene-sized DNA molecules of the macronuclei in three species of hypotrichs: size distributions and absence of nicks. DNA of ciliated protozoa. VIII.

Macronuclear DNA s from three related hypotrichous ciliated protozoans were compared by agarose gel electrophoresis. Each was shown to be composed of DNA duplexes that yielded a unique pattern of bands overlying a continuous distribution of DNA sizes ranging from approximately 400 bp to approximately 20,000 bp. By EM, the number average molecular sizes for double-stranded DNA were 2,200 bp for Oxytricha sp., 2,514 bp for Stylonychia pustulata and 18,836 bp for Euplotes aediculatus. Contrary to previous reports we present evidence that the macronuclear DNA s in each of these three organisms lack single-stranded interruptions.

Animals↗
PubMed1979

Different Chinese hamster cell lines express a G1 period for different reasons.

Previous studies from our laboratory have shown that the absence of G1(G1-condition) in two lines of Chinese hamster cells is dominant over the presence of G1(G1+condition) in a variety of intraspecific cell hybrids. G1+ mutants or variants cna be isolated from G1- cells following mutagenesis and selection. These G1+ mutants fall into multiple complementation groups based on their abilities to form G1- cell hybrids with one another. This is evidence that different mutants have G1 intervals for different reasons, possibly as the result of deficiencies in functions necessary for G1- cell cycles. In this report we have used cell hybrid analysis to ask whether cells of different, naturally occurring G1+ lines of Chinese hamster are able to complement to produce G1- hybrids. We have found three complementation groups among the four G1+ cell lines examined. Therefore, these lines define three different reasons or bases for the existence of a G1 interval. These results lead us to suggest that multiple requirements must be met for these cells to start the S period, but that failure to fulfill only a single and different requirement is responsible for the presence of a G1 interval in any given cell line.

Animals↗
PubMed1979

Resistance to Ehrlich ascites tumor in a strain of dystrophic mice.

Ehrlich ascites tumor grows in almost any mouse host. We have found a strain of dystrophic mice to be strongly resistant to Ehrlich ascites tumor. Mice of the dystrophic strain survived injection of Ehrlich Ascites tumor cells that had just been passaged through either dystrophic mice or C57BL X DBA/2 F1, (hereafter known as BD2F1) mice. However, most of the dystrophic mice died when they were given injections of Ehrlich ascites tumor cells that had just been passaged through Swiss white mice. All Swiss white and BD2F1 mice died when they were inoculated with Ehrlich ascites tumor cells, regardless of the type of mouse through which the Ehrlich ascites tumor cells had just been passaged. In some dystrophic mice, the tumor grew to a palpabale size and then disappeared. This provided the opportunity to passage the tumor from one dystrophic mouse to another and thereby to demonstrate resistance in the dystrophic mice to tumor that had grown previously in dystrophic mice. There was no detectable difference in the rate of tumor growth in either Swiss white or BD2F1 mice.

Animals↗
PubMed1978

Genetic analysis of the G1 period: isolation of mutants (or variants) with a G1 perior from a Chinese hamster cell line lacking G1.

Cells of the Chinese hamster line V79-8 multiply without a G1 period (i.e., they are G1(-)) and have an average generation time of 9.5 hr. After mutagenesis and selection we have derived five stable mutants (or variants) of this line that have longer generation times. In each case the increase in generation time is due solely to the introduction of a G1 period into the cell cycle, with no measurable effect on S, G2, or M. Fusions among these five G1(+) mutant lines and another presumably nonmutant G1(+) line (V79-743) produce hybrid cells lacking a G1 period in all but one case. These complementation tests define five complementation groups among these six G1(+) cell lines. The six G1(+) lines represent five different causes or bases for the presence of a G1 period. The two G1(+) mutants belonging to complementation group V are temperature sensitive for expression of the G1(+) phenotype (G1 congruent with 0, 4, and 6 hr at 33 degrees , 37 degrees , and 39 degrees , respectively). In all cases the G1(-) state is dominant over the G1(+) state, suggesting that the presence of G1 represents a "deficient" condition. Mutants of this type may be useful in the analysis of the switch from G1(-) to G1(+) that occurs normally in cleaving embryos and in elucidation of the genetic mechanism(s) responsible for the presence of a measurable G1 in most cells.

Cell Cycle↗
PubMed1978

Late S phase cells (Chinese hamster ovary) induce early S phase DNA labeling patterns in G1 phase nuclei.

Cells (Chinese hamster ovary) in G1 phase were fused with cells in late S phase to determine if a cell in late S phase can induce DNA synthesis in the nucleus of a G1 cell and, if so, to determine if the DNA synthesis so induced in a G1 phase nucleus has an autoradiographic pattern characteristic of early or of late S phase synthesis. The results indicate (i) that 89% of the G1 nuclei in late-S/G1 binucleates synthesized DNA, while only 2% of the control unfused G1 cells synthesized DNA, and (ii) that in all late-S/G1 binucleates the G1 nucleus was induced to synthesize early S phase DNA. These results are compatible with the idea that a cytoplasmically transmissible factor initiates DNA synthesis but that an intranuclear mechanism defines the temporal order of replication.

Cell Cycle↗
PubMed1976

DNA of ciliated protozoa: DNA sequence diminution during macronuclear development of Oxytricha.

We have measured the reassociation kinetics of DNA from the micronucleus and from the macronucleus of the hypotrichous ciliate Oxytricha. The micronuclear DNA reassociates with at least a two-component reaction, indicating the presence of both repeated and non-repeated sequences. The kinetic complexity of micronuclear non-repeated DNA is in the range of 2 to 15 X 10(11) daltons; the haploid DNA content of the micronucleus is 4 X 10(11) daltons (0.66 pg), measured microspectrophotometrically. The DNA of the macronucleus reassociates as a single second-order reaction, with a kinetic complexity of 3.6 X 10(10) daltons. A comparison of the kinetic complexities of micronuclear and macronuclear DNAs suggest a 5 to 30 fold reduction in DNA sequence complexity during the formation of a macronucleus from a micronucleus. Macronuclear DNA is in pieces with an average molecular weight of 2.1 X 10(6) daltons. Since the kinetic complexity of macronuclear DNA is 3.6 X 10(10) daltons, the macronucleus must contain about 17,000 different kinds of DNA pieces. Each macronucleus contains 3.5 X 10(13) daltons (58 pg) of DNA, indicating that each sequence must be present about 1000 times per macronucleus or 2000 times per cell.

Animals↗
PubMed1974

Reconstruction of mammalian cells from nuclear and cytoplasmic components separated by treatment with cytochalasin B.

Mouse L929 cells were separated into enucleated cytoplasmic components (cytoplasts) and nucleated subcellular fractions (karyoplasts) in the presence of cytochalasin B. Karyoplasts from cells containing tritiated nuclei were fused, using inactivated Sendai virus, to cytoplasts from cells containing large (1.0-mum diameter) latex spheres in the cytoplasm. Mononucleated cells containing radioactive nuclei and large latex spheres in the cytoplasm were observed among the products of the fusion reaction. Some of these cells were in mitotic configurations. The results indicate that cells capable of undergoing mitosis can be reconstructed from the products of cellular enucleation in the presence of cytochalasin B.

Animals↗
PubMed1974

The surface morphology and fine structure of CHO (Chinese hamster ovary) cells following enucleation.

Chinese hamster ovary cells grown in monolayer culture and exposed to cytochalasin B were enucleated by centrifugation. Thereafter, the karyoplasts (the nucleated parts obtained from the bottoms of the centrifuge tubes) and the cytoplasts (the enucleated cytoplasmic parts attached to the coverslips) were allowed to recover and subsequently were examined by scanning and transmission electron microscopy. Microscopy of thin sections revealed that the karyoplasts, limited by an intact plasma membrane, contain an intact nucleus surrounded by a layer of cytoplasm that includes ribosomes, mitochondria, and fragments of the endoplasmic reticulum, but no centrioles or microtubules. The cytoplasts, similarly examined, appear to contain all cytoplasmic organelles and systems, including centrioles and microtubules. The karyoplasts, when replated in fresh medium adhere to the substrate but remain essentially spherical and are incapable of motility. They disintegrate in about 72 hr. The cytoplasts, under identical conditions, recover a shape similar to that of the whole Chinese hamster ovary cell and display some motility. They generally survive not more than 48 hr. It appears that this enucleation procedure consistently separates the nucleus and limited cytoplasm from the centrosphere and microtubule-containing cytoplasts and, furthermore, that the formdetermining and motility mechanisms reside in the cytoplast and function without nuclear participation for the short period of viability.

Animals↗
PubMed1974

Establishment and maintenance of the interferon-induced antiviral state: studies in enucleated cells.

Requirements for the physical presence of the cell's nucleus for the establishment and maintenance of the interferon-induced antiviral state were investigated. Enucleated chicken embryo fibroblasts were obtained by cytochalasin B treatment during centrifugation. The inhibition of vaccinia virus cytoplasmic DNA synthesis, monitored by autoradiography, was used to measure the antiviral activity resulting from interferon treatment. The antiviral state is not established in cells treated with interferon after removal of their nuclei. On the other hand, cells first treated with interferon for 6 or 12 h and then enucleated express the antiviral state. Furthermore, the antiviral state is maintained in enucleated cells for 16 h after enucleation. The antiviral state appears to be more stable in enucleates than in the residual nucleated cells found in the same cultures. Single cells of antiviral populations are found to be either fully permissive or fully restrictive to vaccinia DNA synthesis. The effect of an increasing intracellular multiplicity of infectious virus is to overcome the antiviral cell's block against viral DNA synthesis.

Animals↗