Linked Life Data

1711534

Source:http://linkedlifedata.com/resource/pubmed/id/1711534

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:dateCreated
1991-8-1
pubmed:abstractText
Different agents have been employed to extract the histones and other soluble components from isolated HeLa S3 nuclei during nuclear matrix isolation. We report that 0.2M (NH4)2SO4 is a milder extracting agent than NaCl and LIS (lithium 3,5-diiodosalicylate), on the basis of the apparent preservation of the elaborate fibrogranular network and the residual nucleolus that resemble the in situ structures in whole cells and nuclei, minimal aggregation, and sufficient solubilization of DNA and histones. The importance of intermolecular disulfide bonds, RNA and 37 degrees C stabilization on the structural integrity of the nuclear matrix was examined in detail using sulfydryl alkylating, reducing and oxidizing agents, and RNase A. The data suggest that any disulfides formed during the isolation are not essential for maintaining the structural integrity of the in vitro matrix. However, structural integrity of the matrix is dependent upon RNA and to some degree on disulfides that presumably existed in situ. Sodium tetrathionate and 37 degrees C stabilization of isolated nuclei resulted in nuclear matrices containing an approximately twofold greater amount of protein, RNA and DNA than control preparations. The 37 degrees C incubation, unlike the sodium tetrathionate stabilization, does not appear to induce intermolecular disulfide bond formation. Neither stabilizations resulted in significant differences of the major matrix polypeptide pattern on two-dimensional (2-D) gels stained with Coomassie Blue as compared to that of unstabilized matrix. The major nuclear matrix proteins, other than the lamins, did not react to the Pruss murine monoclonal antibody (IFA) that recognizes all known intermediate filament proteins, suggesting that the internal matrix proteins are not related to the lamins in intermediate filament-like quality.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0021-9533
pubmed:author
pubmed:issnType
Print
pubmed:volume
98 ( Pt 3)
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
281-91
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:1711534-Blotting, Western, pubmed-meshheading:1711534-Cell Fractionation, pubmed-meshheading:1711534-Cell Nucleus, pubmed-meshheading:1711534-DNA, pubmed-meshheading:1711534-Disulfides, pubmed-meshheading:1711534-Electrophoresis, Gel, Two-Dimensional, pubmed-meshheading:1711534-HeLa Cells, pubmed-meshheading:1711534-Histones, pubmed-meshheading:1711534-Humans, pubmed-meshheading:1711534-Hydrogen-Ion Concentration, pubmed-meshheading:1711534-Intermediate Filament Proteins, pubmed-meshheading:1711534-Lamins, pubmed-meshheading:1711534-Microscopy, Electron, pubmed-meshheading:1711534-Nuclear Matrix, pubmed-meshheading:1711534-Nuclear Proteins, pubmed-meshheading:1711534-Nucleolus Organizer Region, pubmed-meshheading:1711534-RNA, pubmed-meshheading:1711534-Salicylic Acids, pubmed-meshheading:1711534-Sodium Chloride, pubmed-meshheading:1711534-Solubility, pubmed-meshheading:1711534-Temperature, pubmed-meshheading:1711534-Tetrathionic Acid
pubmed:year
1991
pubmed:articleTitle
A comprehensive study on the isolation and characterization of the HeLa S3 nuclear matrix.
pubmed:affiliation
Department of Biological Sciences, State University of New York, Buffalo 14260.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.