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Polo-like Kinase I is involved in Invasion through Extracellular Matrix

Description: Polo-like kinase 1, PLK1, has important functions in maintaining genome stability and is involved in regulation of mitosis. PLK1 is up regulated in many invasive carcinomas. We asked whether it may also play a role in acquisition of invasiveness, a crucial step in transition to malignancy. In a model of metaplastic basal-like breast carcinoma progression, we found that PLK1 expression is necessary but not sufficient to induce invasiveness through laminin-rich extracellular matrix. PLK1 mediates invasion via Vimentin and {beta}1 integrin, both of which are necessary. We observed that PLK1 phosphorylates Vimentin on serine 82, which in turn regulates cell surface levels of {beta}1 integrin. We found PLK1 to be also highly expressed in pre-invasive in situ carcinomas of the breast. These results support a role for the involvement of PLK1 in the invasion process and point to this pathway as a potential therapeutic target for pre-invasive and invasive breast carcinoma treatment.
Date: April 2, 2008
Creator: Bissell, Mina J; Rizki, Aylin; Mott, Joni D. & Bissell, Mina J
Partner: UNT Libraries Government Documents Department

A Family of Zinc Finger Proteins Is Required forChromosome-specific Pairing and Synapsis during Meiosis in C.elegans

Description: Homologous chromosome pairing and synapsis are prerequisitefor accurate chromosome segregation during meiosis. Here, we show that afamily of four related C2H2 zinc-finger proteins plays a central role inthese events in C. elegans. These proteins are encoded within a tandemgene cluster. In addition to the X-specific HIM-8 protein, threeadditional paralogs collectively mediate the behavior of the fiveautosomes. Each chromosome relies on a specific member of the family topair and synapse with its homolog. These "ZIM" proteins concentrate atspecial regions called meiotic pairing centers on the correspondingchromosomes. These sites are dispersed along the nuclear envelope duringearly meiotic prophase, suggesting a role analogous to thetelomere-mediated meiotic bouquet in other organisms. To gain insightinto the evolution of these components, wecharacterized homologs in C.briggsae and C. remanei, which revealed changes in copy number of thisgene family within the nematode lineage.
Date: June 7, 2006
Creator: Phillips, Carolyn M. & Dernburg, Abby F.
Partner: UNT Libraries Government Documents Department

Mislocalization of the Drosophila centromere-specific histone CIDpromotes formation of functional ectopic kinetochores

Description: The centromere-specific histone variant CENP-A (CID in Drosophila) is a structural and functional foundation for kinetochore formation and chromosome segregation. Here, we show that overexpressed CID is mislocalized into normally non-centromeric regions in Drosophila tissue culture cells and animals. Analysis of mitoses in living and fixed cells reveals that mitotic delays, anaphase bridges, chromosome fragmentation, and cell and organismal lethality are all direct consequences of CID mislocalization. In addition, proteins that are normally restricted to endogenous kinetochores assemble at a subset of ectopic CID incorporation regions. The presence of microtubule motors and binding proteins, spindle attachments, and aberrant chromosome morphologies demonstrate that these ectopic kinetochores are functional. We conclude that CID mislocalization promotes formation of ectopic centromeres and multicentric chromosomes, which causes chromosome missegregation, aneuploidy, and growth defects. Thus, CENP-A mislocalization is one possible mechanism for genome instability during cancer progression, as well as centromere plasticity during evolution.
Date: January 30, 2006
Creator: Heun, Patrick; Erhardt, Sylvia; Blower, Michael D.; Weiss,Samara; Skora, Andrew D. & Karpen, Gary H.
Partner: UNT Libraries Government Documents Department

Mitotic Exit Control as an Evolved Complex System

Description: The exit from mitosis is the last critical decision a cell has to make during a division cycle. A complex regulatory system has evolved to evaluate the success of mitotic events and control this decision. Whereas outstanding genetic work in yeast has led to rapid discovery of a large number of interacting genes involved in the control of mitotic exit, it has also become increasingly difficult to comprehend the logic and mechanistic features embedded in the complex molecular network. Our view is that this difficulty stems in part from the attempt to explain mitotic exit control using concepts from traditional top-down engineering design, and that exciting new results from evolutionary engineering design applied to networks and electronic circuits may lend better insights. We focus on four particularly intriguing features of the mitotic exit control system: the two-stepped release of Cdc14; the self-activating nature of Tem1 GTPase; the spatial sensor associated with the spindle pole body; and the extensive redundancy in the mitotic exit network. We attempt to examine these design features from the perspective of evolutionary design and complex system engineering.
Date: April 25, 2005
Creator: Bosl, W & Li, R
Partner: UNT Libraries Government Documents Department

Comparative analyses of gene copy number and mRNA expression in GBM tumors and GBM xenografts

Description: Development of model systems that recapitulate the molecular heterogeneity observed among glioblastoma multiforme (GBM) tumors will expedite the testing of targeted molecular therapeutic strategies for GBM treatment. In this study, we profiled DNA copy number and mRNA expression in 21 independent GBM tumor lines maintained as subcutaneous xenografts (GBMX), and compared GBMX molecular signatures to those observed in GBM clinical specimens derived from the Cancer Genome Atlas (TCGA). The predominant copy number signature in both tumor groups was defined by chromosome-7 gain/chromosome-10 loss, a poor-prognosis genetic signature. We also observed, at frequencies similar to that detected in TCGA GBM tumors, genomic amplification and overexpression of known GBM oncogenes, such as EGFR, MDM2, CDK6, and MYCN, and novel genes, including NUP107, SLC35E3, MMP1, MMP13, and DDX1. The transcriptional signature of GBMX tumors, which was stable over multiple subcutaneous passages, was defined by overexpression of genes involved in M phase, DNA replication, and chromosome organization (MRC) and was highly similar to the poor-prognosis mitosis and cell-cycle module (MCM) in GBM. Assessment of gene expression in TCGA-derived GBMs revealed overexpression of MRC cancer genes AURKB, BIRC5, CCNB1, CCNB2, CDC2, CDK2, and FOXM1, which form a transcriptional network important for G2/M progression and/or checkpoint activation. Our study supports propagation of GBM tumors as subcutaneous xenografts as a useful approach for sustaining key molecular characteristics of patient tumors, and highlights therapeutic opportunities conferred by this GBMX tumor panel for testing targeted therapeutic strategies for GBM treatment.
Date: April 3, 2009
Creator: Hodgson, J. Graeme; Yeh, Ru-Fang; Ray, Amrita; Wang, Nicholas J.; Smirnov, Ivan; Yu, Mamie et al.
Partner: UNT Libraries Government Documents Department


Description: Chemical mapping of proteins and lipids inside a single living cell and at a resolution of a few microns, has been performed using synchroton infrared microspectrometry. Modifications of the chemical distributions upon mitosis and necrosis has been investigated.
Date: July 29, 1997
Partner: UNT Libraries Government Documents Department

Microsporogenesis and Embryogenesis in Quercus

Description: Representative species from two subgenera in the genus Quercus were examined for floral structure and phenology, microsporogenesis, and embryogenesis. The species selected for investigation were: Quercus alba in the Lepidobalanus subgenera, and Quercus coccinea and Quercus ilicifolia from the Erythrobalanus group. Photographs of flowering and photomicrographs of microsporogensis and embryogenesis are used for illustration. The male flowers of the three species are borne on catkins which develop in the scale leaf axils of the current vegetative bud or in separate male buds. Meiosis occurred in the spring at the beginning of bud enlargement; division figures were regular in all the material observed. A haploid chromosome number of 12 was confirmed for the three species. Pollen was shed on May 10, 1962, from trees of Quercus coccinea and Quercus ilicifolia; and on May 26, 1962 from Quercus alba. The female flowers are located in the axils of the new leaves. Seed development requires one growing season in Quercus alba, but two growing seasons are required to mature seed of Quercus coccinea and Quercus ilicifolia. The chronology of embryo development was similar for Quercus coccinea and Quercus ilicifolia; embryo development of Quercus alba was about two weeks behind that of the other two species. Definition of ovule dominance within a seed occurred at the time of early embryo development. Failure of this physiological expression of dominance results in multiseeded acorns. No abnormal embryogenesis per se was observed in relation to multiple embryo development. (auth)
Date: January 1, 1962
Creator: Stairs, G. R.
Partner: UNT Libraries Government Documents Department


Description: The reproductive integrity of single meristematic cells of Tradescantia occidentalis exposed to acute doses of x rays was investigated. The dose response curve was sigmoid and similar to that reported for a variety of mammaliand cell lines having a D/sub O/ of 149 r and and extrapolation number of 1.6. Detailed observations were also made of all forms of chromatid and chromosome aberrations induced after irradiating all stages of the mitotic cycle of these same meristematic cells. Attempts were then made to correlate these two sets of data and to equate loss of genetic information to loss of reproductive integrity. (auth)
Date: January 1, 1962
Creator: Davies, D.R.
Partner: UNT Libraries Government Documents Department


Description: The authors have previously reported that deuterium oxide in the drinking water of either male or female mice produces sterility. An investigation of some of the conditions--with particular reference to time--of deuterium oxide treatment to produce sterile C{sub 57} male mice indicated that the sensitive phase of sperm production centered around the late prophase of meiosis. In some experiments, although D{sub 2}O was almost completely absent during maturation of the sperm, and when the mice mated, these sperm exhibited the effects of their much earlier contact with D{sub 2}O. No viable offspring were obtained from these matings. They concluded that the presence of D{sub 2}O during the late prophase and meiotic divisions interfered with the normal construction or division (or both) of genetic material. It was suggested that changes in the forces, principally hydrogen bonds, in macromolecules affected their structural characteristics and resulted in abnormal division. The objective of the experiments reported here was to determine the phases of embryonic development of the mouse at which the lethal action of deuterium oxide on sperm is manifested. These investigations on embryonic growth initiated by sperm developed in D{sub 2}O have yielded additional evidence that D{sub 2}O severely damages the genetic material of developing sperm, with resulting sterility of the male mouse.
Date: June 1, 1960
Creator: Hughes, Ann M.; Bennett, Edward L. & Calvin, Melvin.
Partner: UNT Libraries Government Documents Department

Activity of the kinesin spindle protein inhibitor ispinesib (SB-715992) in models of breast cancer

Description: Ispinesib (SB-715992) is a potent inhibitor of kinesin spindle protein (KSP), a kinesin motor protein essential for the formation of a bipolar mitotic spindle and cell cycle progression through mitosis. Clinical studies of ispinesib have demonstrated a 9% response rate in patients with locally advanced or metastatic breast cancer, and a favorable safety profile without significant neurotoxicities, gastrointestinal toxicities or hair loss. To better understand the potential of ispinesib in the treatment of breast cancer we explored the activity of ispinesib alone and in combination several therapies approved for the treatment of breast cancer. We measured the ispinesib sensitivity and pharmacodynamic response of breast cancer cell lines representative of various subtypes in vitro and as xenografts in vivo, and tested the ability of ispinesib to enhance the anti-tumor activity of approved therapies. In vitro, ispinesib displayed broad anti-proliferative activity against a panel of 53 breast cell-lines. In vivo, ispinesib produced regressions in each of five breast cancer models, and tumor free survivors in three of these models. The effects of ispinesib treatment on pharmacodynamic markers of mitosis and apoptosis were examined in vitro and in vivo, revealing a greater increase in both mitotic and apoptotic markers in the MDA-MB-468 model than in the less sensitive BT-474 model. In vivo, ispinesib enhanced the anti-tumor activity of trastuzumab, lapatinib, doxorubicin, and capecitabine, and exhibited activity comparable to paclitaxel and ixabepilone. These findings support further clinical exploration of KSP inhibitors for the treatment of breast cancer.
Date: June 10, 2009
Creator: Purcell, James W; Davis, Jefferson; Reddy, Mamatha; Martin, Shamra; Samayoa, Kimberly; Vo, Hung et al.
Partner: UNT Libraries Government Documents Department

A Link between Meiotic Prophase Progression and CrossoverControl

Description: During meiosis, most organisms ensure that homologous chromosomes undergo at least one exchange of DNA, or crossover, to link chromosomes together and accomplish proper segregation. How each chromosome receives a minimum of one crossover is unknown. During early meiosis in Caenorhabditis elegans and many other species, chromosomes adopt a polarized organization within the nucleus, which normally disappears upon completion of homolog synapsis. Mutations that impair synapsis even between a single pair of chromosomes in C. elegans delay this nuclear reorganization. We quantified this delay by developing a classification scheme for discrete stages of meiosis. Immunofluorescence localization of RAD-51 protein revealed that delayed meiotic cells also contained persistent recombination intermediates. Through genetic analysis, we found that this cytological delay in meiotic progression requires double-strand breaks and the function of the crossover-promoting heteroduplex HIM-14 (Msh4) and MSH-5. Failure of X chromosome synapsis also resulted in impaired crossover control on autosomes, which may result from greater numbers and persistence of recombination intermediates in the delayed nuclei. We conclude that maturation of recombination events on chromosomes promotes meiotic progression, and is coupled to the regulation of crossover number and placement. Our results have broad implications for the interpretation of meiotic mutants, as we have shown that asynapsis of a single chromosome pair can exert global effects on meiotic progression and recombination frequency.
Date: July 6, 2005
Creator: Carlton, Peter M.; Farruggio, Alfonso P. & Dernburg, Abby F.
Partner: UNT Libraries Government Documents Department

Development of affinity technology for isolating individual human chromosomes by third strand binding

Description: The overall goal was to explore whether nucleic acid third strands could be used to bind with very high specificity to specific targets within whole genomes. Towards this end conditions had to be found to keep erroneous binding to an absolute minimum. The goal to use third strands (linked to magnetic beads) to ''capture'' large particles such as plasmids, cosmids, and whole chromosomes from complex mixtures was partially met; their use to serve as cytogenetic probes of metaphase chromosomes and to deliver reactive reagents to unique target sites on chromosomes in vivo for the purpose of mutagenizing specific base pairs was fully met; and their use as cytogenetic probes of chromosomal DNA in sections of formalin-fixed, paraffin-embedded tissue has been met since the DOE support was terminated.
Date: June 1, 2003
Creator: Fresco, Jacques R.
Partner: UNT Libraries Government Documents Department

Downregulation of Protein 4.1R impairs centrosome function,bipolar spindle organization and anaphase

Description: Centrosomes nucleate and organize interphase MTs and areinstrumental in the assembly of the mitotic bipolar spindle. Here wereport that two members of the multifunctional protein 4.1 family havedistinct distributions at centrosomes. Protein 4.1R localizes to maturecentrioles whereas 4.1G is a component of the pericentriolar matrixsurrounding centrioles. To selectively probe 4.1R function, we used RNAinterference-mediated depletion of 4.1R without decreasing 4.1Gexpression. 4.1R downregulation reduces MT anchoring and organization atinterphase and impairs centrosome separation during prometaphase.Metaphase chromosomes fail to properly condense/align and spindleorganization is aberrant. Notably 4.1R depletion causes mislocalizationof its binding partner NuMA (Nuclear Mitotic Apparatus Protein),essential for spindle pole focusing, and disrupts ninein. Duringanaphase/telophase, 4.1R-depleted cells have lagging chromosomes andaberrant MT bridges. Our data provide functional evidence that 4.1R makescrucial contributions to centrosome integrity and to mitotic spindlestructure enabling mitosis and anaphase to proceed with the coordinatedprecision required to avoid pathological events.
Date: March 17, 2006
Creator: Spence, Jeffrey R.; Go, Minjoung M.; Bahmanyar, S.; Barth,A.I.M. & Krauss, Sharon Wald
Partner: UNT Libraries Government Documents Department

Attenuation of G{sub 2} cell cycle checkpoint control in human tumor cells is associated with increased frequencies of unrejoined chromosome breaks but not increased cytotoxicity following radiation exposure

Description: The contribution of G{sub 2} cell cycle checkpoint control to ionizing radiation responses was examined in ten human tumor cell lines. Most of the delay in cell cycle progression seen in the first cell cycle following radiation exposure was due to blocks in G{sub 2} and there were large cell line-to-cell line variations in the length of the G{sub 2} block. Longer delays were seen in cell lines that had mutations in p53. There was a highly significant inverse correlation between the length of G{sub 2} delay and the frequency of unrejoined chromosome breaks seen as chromosome terminal deletions in mitosis, and observation that supports the hypothesis that the signal for G{sub 2} delay in mammalian cells is an unrejoined chromosome break. There were also an inverse correlation between the length of G{sub 2} delay and the level of chromosome aneuploidy in each cell line, suggesting that the G{sub 2} and mitotic spindel checkpoints may be linked to each other. Attenuation in G{sub 2} checkpoint control was not associated with alterations in either the frequency of induced chromosome rearrangements or cell survival following radiation exposure suggesting that chromosome rearrangements, the major radiation-induced lethal lesion in tumor cells, form before cells enters G{sub 2}. Thus, agents that act solely to override G{sub 2} arrest should produce little radiosensitization in human tumor cells.
Date: August 1997
Creator: Schwartz, J. L.; Cowan, J. & Grdina, D. J.
Partner: UNT Libraries Government Documents Department

Chronic and Acute Radiation of Microsporogenesis and Mature Pollen in Quercus

Description: The production of pollen under conditions of chronic gamma irradiation was investigated for three oak species. Two chronically irradiated areas were studied: a low level (1 to 15r/day) area where trees had received varying amounts of radiation over a period of 11 years, and a second area receiving gamma radiation for about five months previous to the investigation. In the latter study dose levels ranged from lethal (45r/day) to a region of no detectable effect. In both areas pollen abortion showed a significant increase with increasing radiation exposure, although germinable pollen was produced at all survival levels examined. The germinating pollen tube length did not show a significant decrease in the irradiated material examined. In addition to cytological effects there was a marked deiny in floral phenology for both areas. Acute irradiation of male flower buds at different stages of meiosis, and of mature pollen were reported. The radiosensitivity of microsporogenesis was evaluated by cytological scoring at anaphase I, and by pollen abortion, germination, and tube lengih. Both the number of chromosome fragments/100 cells scored at anaphase I and pollen abortion showed a linear increase with an increase in radiation exposure. Pollen germination and tube length were less effected by radiation (based on a percent of unaborted grains). It was suggested that a range of 1 kr to 4 kr will be appropriate for irradiating male flower buds of oak to be utilized in a mutation breeding program. Contingent upon additional studies the range of radiation recommended for flower buds is also suggested for the induction of mutations in pollen. Pollen was found to be highly resistant to radiation when evaluated by germination and tube growth studies. No effect was found with irradiation of 100 kr; at 300 kr both germination and tube lengths were depressed. At these levels it ...
Date: January 1, 1963
Creator: Stairs, G. R.
Partner: UNT Libraries Government Documents Department

Stem-loop structures of the repetitive DNA sequences located at human centromeres

Description: The presence of the highly conserved repetitive DNA sequences in the human centromeres argues for a special role of these sequences in their biological functions - most likely achieved by the formation of unusual structures. This prompted us to carry out quantitative one- and two-dimensional nuclear magnetic resonance (lD/2D NMR) spectroscopy to determine the structural properties of the human centromeric repeats, d(AATGG){sub n.d}(CCATT){sub n}. The studies on centromeric DNAs reveal that the complementary sequence, d(AATGG){sub n.d}(CCATT){sub n}, adopts the usual Watson-Crick B-DNA duplex and the pyrimidine-rich d(CCATT){sub n} strand is essentially a random coil. However, the purine-rich d(AATGG){sub n} strand is shown to adopt unusual stem-loop structures for repeat lengths, n=2,3,4, and 6. In addition to normal Watson-Crick A{center_dot}T pairs, the stem-loop structures are stabilized by mismatch A{center_dot}G and G{center_dot}G pairs in the stem and G-G-A stacking in the loop. Stem-loop structures of d(AATGG)n are independently verified by gel electrophoresis and nuclease digestion studies. Thermal melting studies show that the DNA repeats, d(AATGG){sub n}, are as stable as the corresponding Watson-Crick duplex d(AATGG){sub n.d}(CCATT){sub n}. Therefore, the sequence d(AATGG){sub n} can, indeed, nucleate a stem-loop structure at little free-energy cost and if, during mitosis, they are located on the chromosome surface they can provide specific recognition sites for kinetochore function.
Date: September 1, 1993
Creator: Gupta, G.; Garcia, A. E.; Ratliff, R.; Moyzis, R. K.; Catasti, P.; Hong, Lin et al.
Partner: UNT Libraries Government Documents Department

Programmed Cell Death During Female Gametophyte Development

Description: Endosperm is a storage tissue in the angiosperm seed that is important both biologically and agriculturally. Endosperm is biologically important because it provides nutrients to the embryo during seed development and agriculturally important because it is a significant source of food, feed, and industrial raw materials. Approximately two-thirds of human calories are derived from endosperm, either directly or indirectly through animal feed. Furthermore, endosperm is used as a raw material for numerous industrial products including ethanol. A major event in endosperm development is the transition between the syncytial phase, during which the endosperm nuclei undergo many rounds of mitosis without cytokinesis, and the cellularized phase, during which cell walls form around the endosperm nuclei. Understanding how the syncytial-cellular transition is regulated is agriculturally important because it influences seed size, seed sink strength, and grain weight. However, the molecular processes controlling this transition are not understood. This project led to the identification of the AGL62 gene that regulates the syncytial-cellular transition during endosperm development. AGL62 is expressed during the syncytial phase and suppresses endosperm cellularization during this period. AGL62 most likely does so by suppressing the expression of genes required for cellularization. At the end of the syncytial phase, the FIS PcG complex suppresses AGL62 expression, which allows expression of the cellularization genes and triggers the initiation of the cellularized phase. Endosperm arises following fertilization of the central cell within the female gametophyte. This project also led to the identification of the AGL80 gene that is required for development of the central cell into the endosperm. Within the ovule and seed, AGL80 is expressed exclusively in the central cell and uncellularized endosperm. AGL80 is required for expression of several central cell-expressed genes, including DEMETER and DD46. Together, these data suggest that AGL80 functions as a transcription factor within the central cell ...
Date: September 15, 2004
Creator: Drews, Gary, N.
Partner: UNT Libraries Government Documents Department

Induction of Genomic Instability In Vivo by Low Doses of 137Cs gamma rays

Description: The overall goal of this project is to determine if low doses (below or equal to the level traditionally requiring human radiation protection, i.e. less than or equal to 10 cGy) of low LET radiation can induce genomic instability. The magnitude of genomic instability was measured as delayed chromosome instability in bone marrow cells of exposed mice with different levels of endogenous DNA-dependent protein kinase catalytic subunit (DNA-PKcs) activity, i.e. high (C57BL/6J mice), intermediate (BALB/cJ mice), and extremely low (Scid mice). In addition, at early time points (1 and 4 hrs) following irradiation, levels of activation of nuclear factor-kappa B (NF-{kappa}B), a transcription factor known to be involved in regulating the expression of genes responsible for cell protection following stimuli, were measured in these cells. Bone marrow cells were collected at different times following irradiation, i.e. 1 hr, 4 hrs, 1 month, and 6 months. A total of five mice per dose per strain were sacrificed at each time point for sample collection. As a result, a total of 80 mice from each strain were used. The frequency and the type of metaphase chromosome aberrations in bone marrow cells collected from exposed mice at different times following irradiation were used as markers for radiation-induced genomic instability. A three-color fluorescence in situ hybridization (FISH) protocol for mouse chromosomes 1, 2, and 3 was used for the analysis of delayed stable chromosomal aberrations in metaphase cells. All other visible chromatid-type aberrations and gross structural abnormalities involving non-painted chromosomes were also evaluated on the same metaphase cells used for scoring the stable chromosomal aberrations of painted chromosomes. Levels of nuclear factor-kappa B (NF-{kappa}B) activation were also determined in cells at 1 and 4 hrs following irradiation (indicative of early responses).
Date: January 6, 2006
Creator: Rithidech, Kanokporn; Simon, Sanford, R. & Whorton, Elbert, B.
Partner: UNT Libraries Government Documents Department