scholarly journals Specific DNA sequences associated with the nuclear matrix in synchronized mouse 3T3 cells.

1983 ◽  
Vol 80 (22) ◽  
pp. 6887-6891 ◽  
Author(s):  
G. I. Goldberg ◽  
I. Collier ◽  
A. Cassel
Keyword(s):  
Dna Sequences ◽  
Nuclear Matrix ◽  
3T3 Cells

Oncogenes in Laryngeal Cancer: Serial Passage of Transformed Cellular DNA

1985 ◽  
Vol 93 (3) ◽  
pp. 346-350 ◽  
Author(s):  
William H. Friedman ◽  
Barry N. Rosenblum ◽  
Paul Loewenstein ◽  
Helen Thornton ◽  
George Katsantonis ◽  
...  
Keyword(s):  
Laryngeal Cancer ◽  
Dna Sequences ◽  
Squamous Cell Cancer ◽  
Cell Cancer ◽  
Serial Passage ◽  
3T3 Cells ◽  
Human Dna ◽  
Cellular Dna ◽  
Nih 3T3 ◽  
Narrow Bands

DNA originally extracted from squamous cell cancer of the larynx has been serially passaged through transformed populations of NIH/3T3 mouse fibroblasts. The transformed foci were then harvested, cloned to volume, and incubated with a fresh population of NIH/3T3 cells in a second passage. Transforming efficiencies were enhanced by serial passage. In addition, Southern Blot analysis of the transformed foci revealed hybridization between transformant DNA and human probe DNA from the Alu family of conserved human DNA sequences. In the first passage this hybridization took the form of diffuse homology throughout the entire molecular weight distribution. The second-passage DNA showed “narrow bands” indicating the possibility that an oncogene has been identified in laryngeal cancer and that serial passage has eliminated contaminating human sequences. Repetitive transfection in third- and fourth-passage studies is now being completed.

DNA Sequences Nonrandomly Distributed in Respect to Rat Liver Nuclear Matrix

1990 ◽  
pp. 333-336
Author(s):  
J. Rzeszowska-Wolny ◽  
J. Lanuszewska ◽  
J. Rogolinski
Keyword(s):  
Rat Liver ◽  
Dna Sequences ◽  
Nuclear Matrix

scholarly journals DNA and proteins of the nuclear matrix are the main targets of benzo[a]pyrene's action in rat hepatocytes.

1993 ◽  
Vol 40 (4) ◽  
pp. 559-562 ◽  
Author(s):  
P Widłak ◽  
J Rzeszowska-Wolny
Keyword(s):  
Molecular Weight ◽  
Dna Adducts ◽  
Dna Sequences ◽  
Nuclear Matrix ◽  
Rat Hepatocytes ◽  
The Other ◽  
Matrix Proteins ◽  
Nuclear Matrix Proteins ◽  
Cultured Rat Hepatocytes ◽  
High Molecular Weight Proteins

The binding of [14C]benzo[a]pyrene (B[a]P) to DNA and proteins in total nuclei and subnuclear fractions of cultured rat hepatocytes was compared. The main targets of B[a]P were non-histone high molecular weight proteins of the nuclear matrix and DNA sequences attached to this structure. Following 24 h exposure to B[a]P the amounts of adducts in the nuclear matrix DNA and proteins were twice as high as in total nuclei. After withdrawal of the carcinogen containing medium the level of B[a]P-induced adducts gradually decreased but always remained the highest in the nuclear matrix proteins. Removal of adducts from the nuclear matrix DNA was more efficient than from the other DNA fractions, and 72 h after exposure to the carcinogen the level of DNA adducts in this fraction was similar to that in total nuclei.

scholarly journals Boundary Element-Associated Factor 32B Connects Chromatin Domains to the Nuclear Matrix

2007 ◽  
Vol 27 (13) ◽  
pp. 4796-4806 ◽  
Author(s):  
Rashmi U. Pathak ◽  
Nandini Rangaraj ◽  
Satish Kallappagoudar ◽  
Krishnaveni Mishra ◽  
Rakesh K. Mishra
Keyword(s):  
Boundary Element ◽  
Dna Sequences ◽  
Nuclear Matrix ◽  
Domain Boundary ◽  
Coiled Coil ◽  
Boundary Elements ◽  
Structural Basis ◽  
Chromatin Domain ◽  
Eukaryotic Genomes

ABSTRACT Chromatin domain boundary elements demarcate independently regulated domains of eukaryotic genomes. While a few such boundary sequences have been studied in detail, only a small number of proteins that interact with them have been identified. One such protein is the boundary element-associated factor (BEAF), which binds to the scs′ boundary element of Drosophila melanogaster. It is not clear, however, how boundary elements function. In this report we show that BEAF is associated with the nuclear matrix and map the domain required for matrix association to the middle region of the protein. This region contains a predicted coiled-coil domain with several potential sites for posttranslational modification. We demonstrate that the DNA sequences that bind to BEAF in vivo are also associated with the nuclear matrix and colocalize with BEAF. These results suggest that boundary elements may function by tethering chromatin to nuclear architectural components and thereby provide a structural basis for compartmentalization of the genome into functionally independent domains.

scholarly journals NIH/3T3 cells transfected with human tumor DNA containing activated ras oncogenes express the metastatic phenotype in nude mice.

1985 ◽  
Vol 5 (1) ◽  
pp. 259-262 ◽  
Author(s):  
U P Thorgeirsson ◽  
T Turpeenniemi-Hujanen ◽  
J E Williams ◽  
E H Westin ◽  
C A Heilman ◽  
...  
Keyword(s):  
Nude Mice ◽  
Dna Sequences ◽  
Immune Cell ◽  
Human Tumor ◽  
3T3 Cells ◽  
Metastatic Phenotype ◽  
Ras Oncogenes ◽  
Nih 3T3 ◽  
Tumor Dna ◽  
Nih 3T3 Cells

NIH/3T3 cells transfected with DNA from malignant human tumors produced experimental and spontaneous metastases in nude mice. In contrast, parent or spontaneously transformed NIH/3T3 cells failed to metastasize. The transfected clones contained either activated c-Harvey-ras or N-ras oncogenes. A representative clone (T71-17SA2) which was used to assess selected cellular and host factors relevant to the metastatic process produced lung metastases in 100% of the NIH nude mice recipients, secreted augmented levels of type IV collagenase, and invaded human amnion basement membrane in vitro. Expression of the metastatic phenotype was not related to decreased sensitivity to natural killer cells or macrophage-mediated cytotoxicity. Analysis of the cellular DNA from the T71-17SA2 transfectant and its corresponding metastases, both of which contained activated N-ras oncogenes, revealed a twofold increase in the N-ras-specific DNA sequences in the metastatic cells. Thus, transfection with human tumor DNA containing activated ras oncogenes can induce the complete metastatic phenotype in NIH/3T3 cells by a mechanism apparently unrelated to immune cell killing.

scholarly journals Inactivation of 14-3-3ς Influences Telomere Behavior and Ionizing Radiation-Induced Chromosomal Instability

2000 ◽  
Vol 20 (20) ◽  
pp. 7764-7772 ◽  
Author(s):  
Sonu Dhar ◽  
Jeremy A. Squire ◽  
M. Prakash Hande ◽  
Raymund J. Wellinger ◽  
Tej K. Pandita
Keyword(s):  
Dna Sequences ◽  
Nuclear Matrix ◽  
Mammalian Cells ◽  
Eukaryotic Cell ◽  
Checkpoint Control ◽  
Aberrant Chromosome ◽  
M Phase ◽  
Chromosomal Breaks ◽  
Radiation Induced

ABSTRACT Telomeres are complexes of repetitive DNA sequences and proteins constituting the ends of linear eukaryotic chromosomes. While these structures are thought to be associated with the nuclear matrix, they appear to be released from this matrix at the time when the cells exit from G2 and enter M phase. Checkpoints maintain the order and fidelity of the eukaryotic cell cycle, and defects in checkpoints contribute to genetic instability and cancer. The 14-3-3ς gene has been reported to be a checkpoint control gene, since it promotes G2 arrest following DNA damage. Here we demonstrate that inactivation of this gene influences genome integrity and cell survival. Analyses of chromosomes at metaphase showed frequent losses of telomeric repeat sequences, enhanced frequencies of chromosome end-to-end associations, and terminal nonreciprocal translocations in 14-3-3ς−/− cells. These phenotypes correlated with a reduction in the amount of G-strand overhangs at the telomeres and an altered nuclear matrix association of telomeres in these cells. Since the p53-mediated G1 checkpoint is operative in these cells, the chromosomal aberrations observed occurred preferentially in G2 after irradiation with gamma rays, corroborating the role of the 14-3-3ς protein in G2/M progression. The results also indicate that even in untreated cycling cells, occasional chromosomal breaks or telomere-telomere fusions trigger a G2 checkpoint arrest followed by repair of these aberrant chromosome structures before entering M phase. Since 14-3-3ς−/− cells are defective in maintaining G2 arrest, they enter M phase without repair of the aberrant chromosome structures and undergo cell death during mitosis. Thus, our studies provide evidence for the correlation among a dysfunctional G2/M checkpoint control, genomic instability, and loss of telomeres in mammalian cells.

scholarly journals Atm Inactivation Results in Aberrant Telomere Clustering during Meiotic Prophase

1999 ◽  
Vol 19 (7) ◽  
pp. 5096-5105 ◽  
Author(s):  
Tej K. Pandita ◽  
Christoph H. Westphal ◽  
Melanie Anger ◽  
Satin G. Sawant ◽  
Charles R. Geard ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Nuclear Matrix ◽  
Meiotic Prophase ◽  
Nuclear Periphery ◽  
Early Prophase ◽  
Telomeric Dna ◽  
Gonadal Atrophy ◽  
Significant Difference ◽  
Atm Protein ◽  
And Control

ABSTRACT A-T (ataxia telangiectasia) individuals frequently display gonadal atrophy, and Atm −/− mice show spermatogenic failure due to arrest at prophase of meiosis I. Chromosomal movements take place during meiotic prophase, with telomeres congregating on the nuclear envelope to transiently form a cluster during the leptotene/zygotene transition (bouquet arrangement). Since the ATM protein has been implicated in telomere metabolism of somatic cells, we have set out to investigate the effects of Atm inactivation on meiotic telomere behavior. Fluorescent in situ hybridization and synaptonemal complex (SC) immunostaining of structurally preserved spermatocytes I revealed that telomere clustering occurs aberrantly inAtm −/− mice. Numerous spermatocytes ofAtm −/− mice displayed locally accumulated telomeres with stretches of SC near the clustered chromosome ends. This contrasted with spermatogenesis of normal mice, where only a few leptotene/zygotene spermatocytes I with clustered telomeres were detected. Pachytene nuclei, which were much more abundant in normal mice, displayed telomeres scattered over the nuclear periphery. It appears that the timing and occurrence of chromosome polarization is altered in Atm −/− mice. When we examined telomere-nuclear matrix interactions in spermatocytes I, a significant difference was observed in the ratio of soluble versus matrix-associated telomeric DNA sequences between meiocytes ofAtm −/− and control mice. We propose that the severe disruption of spermatogenesis during early prophase I in the absence of functional Atm may be partly due to altered interactions of telomeres with the nuclear matrix and distorted meiotic telomere clustering.

scholarly journals Identification of ErbB3-stimulated genes using modified representational difference analysis

1997 ◽  
Vol 323 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Carl F. EDMAN ◽  
Sally A. PRIGENT ◽  
Andrea SCHIPPER ◽  
James R. FERAMISCO
Keyword(s):  
Dna Sequences ◽  
Tyrosine Kinases ◽  
Growth Factor Receptor ◽  
Representational Difference Analysis ◽  
3T3 Cells ◽  
Difference Analysis ◽  
Total Rna ◽  
Egfr Family ◽  
Nih 3T3 ◽  
Nih 3T3 Cells

The epidermal growth factor receptor (EGFR) family of tyrosine kinases is involved in the growth of normal and tumour cells. The specific contribution of each of the four family members to these processes remains unclear. In the present study we have used a PCR-based subtractive approach to identify differences in messages induced in response to activation of ErbB3 and EGFR. The approach described is a modification of the representational difference analysis technique adapted for analysis of cDNA, which we have modified to permit identification of differential gene expression using as little as 20 μg of total RNA as the starting material. The mRNA obtained from EGF-stimulated NIH-3T3 cells expressing chimaeric EGFR-ErbB3 receptors provided the tester amplicons (small PCR-amplified fragments) which were subtracted against driver amplicons derived from unstimulated NIH-3T3 cells expressing the EGFR-ErbB3 chimaera or EGF-stimulated NIH-3T3 cells overexpressing the EGFR. A total of 22 different clones were isolated, 90% of which showed increased expression in the tester amplicons. Six of these, corresponding to known DNA sequences, were selected for further Northern blot analysis against total RNA prepared from the starting cell lines. Of these, the gene encoding the protein dlk (or a closely related protein, Pref-1) was identified as being regulated by ErbB3 but not by the EGFR. Other genes appeared to be elevated by both ErbB3 and EGFR, including those encoding c-jun, Ret finger protein (RFP), neuroleukin and amyloid protein precursor. One gene product, TIS11, was identified as being regulated by EGFR but not by ErbB3.

Phosphoinositidase C Isoforms Are Specifically Localized in the Nuclear Matrix and Cytoskeleton of Swiss 3T3 Cells

1993 ◽  
Vol 208 (1) ◽  
pp. 257-269 ◽  
Author(s):  
Nicoletta Zini ◽  
Alberto M. Martelli ◽  
Lucio Cocco ◽  
Francesco A. Manzoli ◽  
Nadir M. Maraldi
Keyword(s):  
Nuclear Matrix ◽  
3T3 Cells ◽  
Swiss 3T3
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