scholarly journals MspI8, the repetitive sequence specifically interacting with nuclear matrix of rat testis cells.

1994 ◽  
Vol 41 (4) ◽  
pp. 459-466 ◽  
Author(s):  
J Rzeszowska-Wolny ◽  
J Rogoliński
Keyword(s):  
Nuclear Matrix ◽  
Gel Electrophoresis ◽  
Matrix Protein ◽  
Repetitive Sequences ◽  
Matrix Proteins ◽  
Nuclear Matrix Proteins ◽  
Rat Testis ◽  
The Matrix ◽  
Matrix Bound

The nuclear matrix bound DNA fraction of rat testis showed enrichment in repetitive sequences found in the 450 bp band after gel electrophoresis of the MspI digested rat DNA. DNA fragments isolated from this band were cloned. DNA of the clone pMspI8 showed homology to some representatives of rat LINE sequence family, and complexed in vitro more efficiently with testes nuclear matrix proteins than with yeast ARS1 sequence containing the matrix association region (MAR) or DNA from an other clone, MspI19. Western blot analysis showed that MspI8 sequence interacts with testes matrix protein of about 120 kDa.

scholarly journals Interactions of rat repetitive sequence MspI8 with nuclear matrix proteins during spermatogenesis.

1996 ◽  
Vol 43 (2) ◽  
pp. 319-324
Author(s):  
J Rogoliński ◽  
P Widłak ◽  
J Rzeszowska-Wolny
Keyword(s):  
Nuclear Matrix ◽  
Blot Analysis ◽  
Repetitive Sequence ◽  
Matrix Proteins ◽  
Velocity Sedimentation ◽  
Nuclear Matrix Proteins ◽  
Rat Testis ◽  
Sedimentation Technique

Using the Southwestern blot analysis we have studied the interactions between rat repetitive sequence MspI8 and the nuclear matrix proteins of rat testis cells. Starting from 2 weeks the young to adult animals showed differences in type of testis nuclear matrix proteins recognizing the MspI8 sequence. The same sets of nuclear matrix proteins were detected in some fractions enriched in spermatocytes and spermatides and obtained after fractionation of testis cells of adult animals by the velocity sedimentation technique.

scholarly journals Ultrastructural localization of nuclear matrix proteins in HeLa cells using silver-enhanced ultra-small gold probes.

1991 ◽  
Vol 39 (8) ◽  
pp. 1035-1045 ◽  
Author(s):  
A de Graaf ◽  
P M van Bergen en Henegouwen ◽  
A M Meijne ◽  
R van Driel ◽  
A J Verkleij
Keyword(s):  
Hela Cells ◽  
Nuclear Matrix ◽  
Matrix Protein ◽  
Ultrastructural Localization ◽  
Matrix Proteins ◽  
Cell Permeabilization ◽  
Immunogold Staining ◽  
Thin Sections ◽  
Nuclear Matrix Proteins ◽  
Dna And Rna

We describe a method for immunogold staining of nuclear matrix proteins using ultra-small gold particles. The nuclear matrix of HeLa cells is obtained by two fractionation steps: (a) cell permeabilization with Triton X-100 to isolate the cytoskeleton, and (b) nuclease digestion followed by an incubation in 0.25 M ammonium sulfate to isolate the nuclear matrix. To prevent redistribution of internal matrix proteins during nuclear matrix preparation, pre-fixation with 0.1% acrolein was performed. Under this condition up to 80% of protein and 90% of DNA and RNA could be removed on nuclear matrix isolation, without redistribution of internal nuclear matrix proteins. For immunogold labeling, 1-nm gold probes appeared to be required to obtain optimal penetration into the nucleus. These particles can be visualized after silver enhancement. After gold labeling the matrices are stained, embedded in Epon, and ultra-thin sections are prepared for examination in the electron microscope. The applicability of this method is examplified by the localization of a 125 KD internal nuclear matrix protein and the lamins A and C in nuclear matrix preparations of HeLa cells.

scholarly journals Developmental association of the beta-galactoside-binding protein galectin-1 with the nuclear matrix of rat calvarial osteoblasts

1998 ◽  
Vol 111 (20) ◽  
pp. 3035-3043 ◽  
Author(s):  
J.Y. Choi ◽  
A.J. van Wijnen ◽  
F. Aslam ◽  
J.D. Leszyk ◽  
J.L. Stein ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Osteoblast Differentiation ◽  
Matrix Protein ◽  
Binding Protein ◽  
Protein Composition ◽  
Matrix Proteins ◽  
Specific Gene ◽  
Two Dimensional Gel Electrophoresis ◽  
Nuclear Matrix Proteins ◽  
Differential Retention

The protein composition of the nuclear matrix changes significantly as the osteoblast matures from a proliferating pre-osteoblast to an osteocyte embedded in a mineralized matrix. These matrix protein are the result of developmental stage-specific gene expression during osteoblast differentiation. To isolate nuclear matrix proteins unique to the bone phenotype we analyzed nuclear matrix preparations from cultures of rat calvarial osteoblasts by high resolution two-dimensional gel electrophoresis at two different stages: proliferation (day 3) and differentiation (day 18, mineralized). We characterized one protein (14 kDa; pI 5.0), that was detectable only in the nuclear matrix of differentiated osteoblasts. By mass spectrometry and microsequencing, this protein was identified as the beta -galactoside-binding protein galectin-1. Both immunofluorescence staining of nuclear matrix preparations with the galectin-1 antibody and western blot analysis of subcellular fractions confirmed that galectin-1 is only associated with the nuclear matrix in differentiated osteoblasts as the result of differential retention. Galectin-1 protein and mRNA are present throughout osteoblast differentiation. Galectin-1 is present in the cytoplasmic and nuclear fractions in both proliferating and differentiated osteoblasts. However, its only stable binding is to the nuclear matrix of the differentiated osteoblast; but, in proliferating osteoblasts, galectin-1 is not retained in the nuclear matrix. Taken together, our results suggest that developmental association of galectin-1 with the nuclear matrix reflects differential subnuclear binding of galectin-1 during osteoblast differentiation.

DNA-binding properties of nuclear matrix proteins

1978 ◽  
Vol 34 (1) ◽  
pp. 233-246 ◽  
Author(s):  
D.E. Comings ◽  
A.S. Wallack
Keyword(s):  
Dna Binding ◽  
Nuclear Matrix ◽  
Matrix Proteins ◽  
Binding Properties ◽  
E Coli ◽  
Nuclear Matrix Proteins ◽  
The Matrix ◽  
Filter Assay ◽  
Dna Binding Properties ◽  
Competition Assays

Mouse nuclear matrix proteins, examined by a filter assay, were found to bind to DNA. There was no preference for homologous mouse compared to heterologous E. coli DNA. Competition assays showed a preference for AT-rich DNA and of the 4 single-stranded homopolymers there was a preference for poly(dT). These observations are consistent with the possibility that the matrix may play a role in the formation of AT-rich chromomeres (G-bands).

Noncovalent Binding of Poly(ADP-Ribose) to Nuclear Matrix Proteins: Developmental Changes and Tissue Specificity

2000 ◽  
Vol 381 (11) ◽  
pp. 1047-1053 ◽  
Author(s):  
M. Malanga ◽  
B. Farina
Keyword(s):  
Nuclear Matrix ◽  
Noncovalent Interactions ◽  
Specific Protein ◽  
Noncovalent Interaction ◽  
Matrix Proteins ◽  
Nuclear Proteins ◽  
Rat Tissues ◽  
Nuclear Matrix Proteins

Abstract Poly(ADP-ribose) is a nuclear polynucleotide involved in the regulation of chromatin functions via covalent and/or noncovalent modification of nuclear proteins. Using a binding assay on protein blots, we searched for poly(ADP-ribose) binding proteins in nuclear matrices from testes of differently aged rats as well as from various adult rat tissues (brain, liver, spleen). We found that nuclear matrix proteins represent a significant subset of the nuclear proteins that can establish noncovalent interactions with poly(ADP-ribose). The profiles of poly(ADP-ribose) binding nuclear matrix proteins appeared to be tissue-specific and changed during postnatal development in the testis. The isolation and analysis of endogenous poly (ADP-ribose) from rat testes showed that the ADP-ribose polymers that bind nuclear matrix proteins in vitro are also present under physiologic conditions in vivo. These results further substantiate the possibility that poly(ADP-ribose) may affect chromatin functions through noncovalent interaction with specific protein targets, including nuclear matrix components.

Proteins From Oyster Shell: Biomineralization Regulators and Commercial Polymer Analogs

1999 ◽  
Vol 599 ◽  
Author(s):  
A. P. Wheeler ◽  
C. S. Sikes
Keyword(s):  
Crystal Growth ◽  
Aspartic Acid ◽  
Large Scale ◽  
Matrix Protein ◽  
Gene Families ◽  
Eastern Oyster ◽  
Matrix Proteins ◽  
The Matrix

AbstractMolluscan shell is a composite made up of μm-sized CaCO3 crystals and an organic phase (matrix). This report outlines our studies on the structure and activities of matrix proteins isolated from the inner calcite layer of shell of the Eastern oyster, including their cellular origin and structure and their relationship to the crystalline mineral phase. In addition, we present results of the synthesis and commercialization of polypeptide polymers which are based on the structure and activities of the oyster proteins. Extracted shell proteins are polyanionic and range in size from relatively small soluble forms to those which are crosslinked and insoluble. The soluble forms are capable of adsorbing to calcite in vitro and in the process changing its growth habit and acting as threshold growth inhibitors. Their function in vivo is not understood, but they may serve to control shell crystal morphology. The insoluble protein forms gels readily and may serve to provide resiliency to the shell and, from in vitro and in situ observations, appears to serve as a site for nucleation of crystals. However, from studies in vitro, these gels do not lower the energy of activation for nucleation, as previously expected. Matrix protein aggregates are identifiable by AFM on the surface of crystals, but as such do not serve as nucleation sites for new crystal growth. If the aggregates are removed, then ectopic crystal growth proceeds readily revealing orientation of the underlying crystals. All the matrix proteins contain domains rich in aspartic acid, are heavily phosphorylated, crossreact in antibody studies and may belong to a limited number of gene families with individuals modified post-synthesis. The proteins are made by a specialized group of cells located primarily some distance from the growing edge of the shell and appear to be assembled into sheets soon after secretion. Soluble anti-scalants and crosslinked insoluble water absorbents have been developed based on the structure and activity of the matrix proteins. These are primarily poly(aspartates) which can be made in large scale via thermal polycondensation of aspartic acid. The soluble forms are commercially used as biodegradable water treatment chemicals among other applications.

In Vitro Heat Exposure Induces a Redistribution of Nuclear Matrix Proteins in Human K562 Erythroleukemia Cells

1994 ◽  
Vol 213 (1) ◽  
pp. 275-285 ◽  
Author(s):  
L.M. Neri ◽  
S. Santi ◽  
R.A. Marugg ◽  
B.M. Riederer ◽  
S. Capitani ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Heat Exposure ◽  
Matrix Proteins ◽  
Nuclear Matrix Proteins ◽  
Erythroleukemia Cells

scholarly journals Identification of distinct messenger RNAs for nuclear lamin C and a putative precursor of nuclear lamin A.

1984 ◽  
Vol 98 (3) ◽  
pp. 980-985 ◽  
Author(s):  
J F Laliberté ◽  
A Dagenais ◽  
M Filion ◽  
V Bibor-Hardy ◽  
R Simard ◽  
...  
Keyword(s):  
Nuclear Matrix ◽  
Gel Electrophoresis ◽  
Lamin A ◽  
Messenger Rnas ◽  
Two Dimensional Gel Electrophoresis ◽  
Nuclear Matrix Proteins ◽  
Nuclear Lamin ◽  
Sds Page ◽  
Putative Precursor

The lamins are the major components of the nuclear matrix and are known as lamins A, B, and C with Mr 72,000, 68,000, and 62,000 when analysed by SDS PAGE. These three polypeptides are very similar, as determined by polypeptide mapping and immunological reactivity. Lamins A and C are so homologous that a precursor-product relationship has been proposed. Using an antiserum against nuclear matrix proteins that specifically immunoprecipitates the three lamins, we examined their synthesis in the rabbit reticulocytes lysate. Four bands of Mr 62,000, 68,000, 70,000, and 74,000 were specifically immunoprecipitated when polysomes or polyadenylated RNA were translated in vitro. By two-dimensional gel electrophoresis, the 68,000- and the 62,000-mol-wt proteins were identified as lamins B and C, respectively, and the 74,000-mol-wt polypeptide had properties of a precursor of lamin A. The mRNAs of lamin C and of the putative precursor of lamin A were completely separated by gel electrophoresis under denaturing conditions, and their respective sizes were determined. These results suggest that lamin A is not a precursor of lamin C.

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