Preferential association of a transcriptionally active gene with the nuclear matrix of rat fibroblasts transformed by a simian-virus-40–pBR322 recombinant plasmid
- 15 April 1990
- journal article
- research article
- Published by Portland Press Ltd. in Biochemical Journal
- Vol. 267 (2), 385-390
- https://doi.org/10.1042/bj2670385
Abstract
To study the relationship between the structural organization and function of the eukaryotic genome, DNA associated with nuclear matrix was analysed by using a transformed rat fibroblast cell line. The nuclear matrices were prepared from the isolated nuclei of pSV1-FR, a rat fibroblast cell line transformed by a pBR322-based recombinant plasmid containing an early gene region, which codes for large T-antigen, of simian virus 40. This transformed cell contained a single copy of the plasmid sequence integrated into the chromosomal DNA of the host cell. The early gene of this plasmid was constitutively expressed, as demonstrated by positive immunofluorescence staining of the cell for large T-antigen and by RNA-blot analysis for its specific mRNA. DNAs were extracted from whole isolated nuclei and nuclear-matrix preparations of the cells, and the relative amounts of the sequence similar to that of the plasmid were compared between these DNA preparations. By employing dot hybridization and Southern-blot analyses we found that the plasmid sequence was more enriched in the DNA extracted from the nuclear matrices than in the DNA extracted from the whole nuclei. When an albumin gene sequence that was not transcribed in this cell line was compared similarly as a control, we found no significant enrichment of this sequence in the DNA associated with the nuclear matrix. Our results strongly support the concept that a transcriptionally active gene is preferentially associated with the nuclear matrix.This publication has 45 references indexed in Scilit:
- Enrichment of selected active human gene sequences in the placental deoxyribonucleic acid fraction associated with tightly bound nonhistone chromosomal proteinsBiochemistry, 1981
- Dynamic association of replicating DNA fragments with the nuclear matrix of regenerating liverExperimental Cell Research, 1981
- Localization of SV40 genes within supercoiled loop domainsNucleic Acids Research, 1980
- Supercoiled loops and eucaryotic DNA replicationCell, 1980
- Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.Proceedings of the National Academy of Sciences, 1980
- Deletions covering the putative promoter region of early mRNAs of simian virus 40 do not abolish T-antigen expression.Proceedings of the National Academy of Sciences of the United States of America, 1980
- Chicken reticulocyte nuclear antigen: its identification and relation to transcriptive activity in erythropoietic cellsBiochemistry, 1980
- Distribution of Active Gene Sequences: A Subset Associated with Tightly Bound Chromosomal ProteinsScience, 1980
- Isolation of biologically active ribonucleic acid from sources enriched in ribonucleaseBiochemistry, 1979
- Simple isolation of DNA hydrophobically complexed with presumed gene regulatory proteins (M3)Biochemistry, 1979