Considerations in the isolation of rat liver nuclear matrix, nuclear envelope, and pore complex lamina

Kaufmann, Scott H.; Coffey, Donald S.; Shaper, Joël H.

doi:10.1016/0014-4827(81)90088-4

Cited by 260 publications

(91 citation statements)

References 68 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This structure has been defined In two ways. Morphologically, the structures remaining after extraction of nuclei with salt to remove most histones and other proteins appear to represent a proteinaceous network of fibers as observed by electron microscopy (3)(4)(5), although the composition and morphology of these structures has engendered considerable controversy (6,7).…”

Section: Introductionmentioning

confidence: 99%

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

1985

View full text Add to dashboard Cite

ABSIRACTUsing the transcriptional modulation afforded by heat shock, we found that the association of active genes with the nuclear matrix was not dependent on their level of transcription. Heat shock genes were matrix associated both before heat shock (when transcription was relatively low), and during heat shock (when transcription was greatly Increased). Conversely, the cytoplasmic actin gene was matrix associated during normal growth conditions (when transcription was high) and during heat shock (when transcription was greatly decreased). Removal of greater than 99.7% of nascent RNA during preparation of the matrices did not affect these findings. Detailed examination of the cytoplasmic actin gene revealed that Its matrix association was apparently mediated by multiple Interactions near the 5' end of the gene.

show abstract

Section: Introductionmentioning

confidence: 99%

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

1985

View full text Add to dashboard Cite

show abstract

“…The nuclear matrix fractions which were somewhat less complex consisted primarily of higher-molecular-weight polypeptides. As noted by others (5,43), abundant quantities of components that are most likely vimentin (molecular weight, 58,000) and actin (molecular weight, 43,000), judging by their relative electrophoretic mobility, were observed in this fraction, as were the characteristic matrix lamins (molecular weight, 62,000 to 69,000) (2,22 T-antigen immunofluorescence in nuclei and nuclear matrices of C6 (A, B) and Cos (C, D) cells. C6 and Cos cells were extracted as described in the text to isolate nuclei (A and C), by lysis of cells in NR buffer, and nuclear matrices (B and D), by treatment of nuclei with DNase I and 2 M NaCl.…”

mentioning

confidence: 52%

“…Several studies have addressed themselves to the properties of the nuclear matrix, identifying its unique components and visualizing, by microscopy, its structural elements (2,4,22). In addition, evidence has been provided suggesting that many processes involved in gene regulation may be performed in association with this structure (8,21,32,36).…”

Section: Discussionmentioning

confidence: 99%

Association of simian virus 40 T antigen with the nuclear matrix of infected and transformed monkey cells.

Covey¹,

Choi²,

Prives³

1984

Mol. Cell. Biol.

View full text Add to dashboard Cite

The subnuclear distribution of simian virus 40 large T antigen within nuclei of transformed Cos and C6 monkey cells was examined. Cos cells express wild-type T antigen but lack viral sequences required for DNA replication, whereas C6 cells contain a functional viral origin but express a replication-defective mutant T antigen which is unable to bind specifically to viral DNA. Discrete subpopulations of T antigen were isolated from the soluble nucleoplasm, chromatin, and nuclear matrix of both cell lines. Although only a small quantity (2 to 12%) of the total nuclear T antigen from Cos cells was associated with the nuclear matrix, a high proportion (25 to 50%) of C6 T antigen was bound to this structure. Results obtained from lytically infected monkey cells showed that early in infection, before viral replication was initiated, a higher proportion (22%) of T antigen was found associated with the nuclear matrix compared with amounts found associated with this structure later in infection (5 to 8%). These results suggest that an increased association of T antigen with this structure is not correlated with viral replication. T antigen isolated from the C6 nuclear matrix was more highly phosphorylated than was soluble C6 T antigen and was capable of binding to the host p53 protein. C6 DNA contains three mutations: two corresponding to N-terminal changes at amino acid positions 30 and 51 and a third located internally at amino acid position 153. By analysis of the subnuclear distribution of T antigen from rat cells transformed by C6 submutant T antigens, it was determined that one or both of the mutations at the NH2 terminus are responsible for the increased quantity of C6 T antigen associated with the nuclear matrix. These results suggest that neither a functional viral DNA replication origin nor the origin binding property of T antigen is required for association of this protein with the nuclear matrix.The simian virus 40 (SV40) A gene product, the large tumor antigen, is required both for viral DNA replication and for oncogenic cell transformation (for a review see reference 47). The existence of T antigen mutants (9,16,33) that lack one or the other of these activities suggests that this protein is multifunctional, with roles in these two processes that are separate and independent. The functional complexity of T antigen is accompanied by a parallel biochemical and immunological heterogeneity. T antigen exists in infected and transformed cells as slowly and rapidly sedimenting forms that vary in their DNA-binding properties, posttranslational modification, and association with the host p53 protein (3,6,12,13,18,28,31,34). Studies with monoclonal antibodies have also provided evidence that T antigen forms immunologically distinct subpopulations (19,20,37). A protein as complicated as this may therefore be expected to exhibit heterogeneity with respect to its association with cellular structures. Although a small proportion of T antigen is associated with the cell membrane (11,41,42,45), the vast majority of the prot...

show abstract

“…In these reports the investigators have taken into consideration the effect on nuclear matrix structure of RNAse A (Kaufmann et al 1981;Belgrader et al 1991), of reducing or oxidative chemicals (Dijkwel and Wenink 1984;Kaufmann and Shaper 1984;Stuurman et al 1992a), and of different extracting agents (Belgrader et al 1991;Eberharter et al 1993). Surprisingly, however, the effect of different concentrations of Mg ++ ions during nuclear preparation to obtain the matrix has never been investigated.…”

Section: Discussionmentioning

confidence: 99%

Different Concentrations of Mg⁺⁺Ions Affect Nuclear Matrix Protein Distribution During Thermal Stabilization of Isolated Nuclei

Neri

Riederer

Valmori

et al. 1997

J Histochem Cytochem.

View full text Add to dashboard Cite

SUMMARYThe nuclear matrix, a proteinaceous network believed to be a scaffolding structure determining higher-order organization of chromatin, is usually prepared from intact nuclei by a series of extraction steps. In most cell types investigated the nuclear matrix does not spontaneously resist these treatments but must be stabilized before the application of extracting agents. Incubation of isolated nuclei at 37C or 42C in buffers containing Mg ++ has been widely employed as stabilizing agent. We have previously demonstrated that heat treatment induces changes in the distribution of three nuclear scaffold proteins in nuclei prepared in the absence of Mg ++ ions. We studied whether different concentrations of Mg ++ (2.0-5 mM) affect the spatial distribution of nuclear matrix proteins in nuclei isolated from K562 erythroleukemia cells and stabilized by heat at either 37C or 42C. Five proteins were studied, two of which were RNA metabolism-related proteins (a 105-kD component of splicing complexes and an RNP component), one a 126-kD constituent of a class of nuclear bodies, and two were components of the inner matrix network. The localization of proteins was determined by immunofluorescent staining and confocal scanning laser microscope. Mg ++ induced significant changes of antigen distribution even at the lowest concentration employed, and these modifications were enhanced in parallel with increase in the concentration of the divalent cation. The different sensitivity to heat stabilization and Mg ++ of these nuclear proteins might reflect a different degree of association with the nuclear scaffold and can be closely related to their functional or structural role.

show abstract

Considerations in the isolation of rat liver nuclear matrix, nuclear envelope, and pore complex lamina

Cited by 260 publications

References 68 publications

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

Association of simian virus 40 T antigen with the nuclear matrix of infected and transformed monkey cells.

Different Concentrations of Mg⁺⁺Ions Affect Nuclear Matrix Protein Distribution During Thermal Stabilization of Isolated Nuclei

Contact Info

Product

Resources

About

Considerations in the isolation of rat liver nuclear matrix, nuclear envelope, and pore complex lamina

Cited by 260 publications

References 68 publications

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

The association of transcribed genes with the nuclear matrix ofDrosophilacells during heat shock

Association of simian virus 40 T antigen with the nuclear matrix of infected and transformed monkey cells.

Different Concentrations of Mg++Ions Affect Nuclear Matrix Protein Distribution During Thermal Stabilization of Isolated Nuclei

Contact Info

Product

Resources

About

Different Concentrations of Mg⁺⁺Ions Affect Nuclear Matrix Protein Distribution During Thermal Stabilization of Isolated Nuclei