Sequence analysis of nuclear matrix associated DNA from rat liver
TL;DR: Using 125I-labeled ribosomal RNA (rRNA) as a probe, it is found that the genes for rRNA are significantly enriched in the residual matrix DNA, indicating that matrix DNA does not represent any specific sequence class.
About: This article is published in Experimental Cell Research.The article was published on 1980-08-01. It has received 71 citations till now. The article focuses on the topics: Nuclear DNA & DNA.
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01 Jan 1985
TL;DR: In this paper, the authors describe the structure of preribosomes and their role in the pre-rRNA precoding process, and discuss the role of RNA polymerases in the precoding.
Abstract: I. Introduction.- II. Ribosomal Genes.- II. 1. Definitions.- II.2. Ribosomal RNA Genes.- II.2.1. Multiplicity.- II.2.2. Chromosomal Location.- II.2.3. Extrachromosomal rRNA Genes.- II.2.4. Organization and Structure.- II.2.4.1. Saccharomyces cerevisiae.- II.2.4.2. Tetrahymena.- II.2.4.3. Drosophila.- II.2.4.4. Xenopus laevis.- II.2.4.5. Higher Plants.- II.2.4.6. Mammalia.- II.2.5. General Features.- II.3. 5 S rRNA Genes.- II.3.1. Number and Chromosomal Location.- II.3.2. Organization and Structure.- II.4. Ribosomal Protein Genes.- II.5. Synopsis.- III. Transcription of Ribosomal Genes.- III. 1. Components of the Transcription Complex.- III. 1.1. RNA Polymerases.- III. 1.2. Nucleolar rDNA and r-Chromatin.- III.2. The Transcription Process >.- III.2.1. Topology of Primary Pre-rRNA.- III.2.2. Morphology of Transcribed rRNA Genes.- III.2.3. Transcribed and Non-Transcribed r-Chromatin.- III.2.4. Primary Transcripts and Primary Pre-rRNA.- III.2.5. Transcription Initiation and Termination.- III.2.5.1. Initiation.- III.2.5.2. Termination.- III.2.6. Transcription in vitro.- III.3. Transcription of 5 S rRNA Genes.- III.4. Transcription of r-Protein Genes.- III.5. Synopsis.- IV. Maturation of Preribosomes.- IV. 1. Structure of Primary Pre-rRNA.- IV. 1.1. Size and Primary Structure.- IV. 1.2. Modifications.- IV. 1.3. Conformation.- IV.2. Pre-rRNA Maturation Pathways.- IV.2.1. General Considerations.- IV.2.2. Common Pattern of Pre-rRNA Maturation.- IV.2.3. Multiplicity of Maturation Pathways.- IV. 2.4. Enzyme Mechanisms.- IV. 3. Preribosomes: Structure and Maturation.- IV. 4. Synopsis.- V. Molecular Architecture of the Nucleolus.- V. 1. Introduction.- V.2. Nucleolus Organizer.- V. 2.1. Chromosomes.- V.2.2. Interphase Nuclei.- V.3. Fibrillar and Granular Components.- V.3.1. The Fibrillar Component.- V.3.2. The Granular Component.- V.4. The Nucleolus and Other Nuclear Structures.- V.4.1. Nucleolus-Associated Chromatin.- V.4.2. The Junction with the Nuclear Envelope.- V.5. The Nucleolar Matrix.- V.6. Macromolecular Constituents.- V.6.1. DNA and RNA.- V. 6.2. Nucleolar Proteins.- V.6.2.1. General.- V.6.2.2. Ag-NOR Protein(s).- V. 6.2.3. Nucleolar Antigens.- V. 7. Outline.- VI. Regulation.- VI. l. General Considerations.- VI.2. Transscriptional Control.- VI. 2.1. Transitions in the State of Expression of rRNA Genes.- VI. 2.1.1. Inactive r-Chromatin.- VI.2.1.2. Potentialy Active and Transcribed rRNA Genes.- VI.2.2. Control of Transcription Rate.- VI.2.2.1. Role of RNA Polymerase I.- VI.2.2.2. Supply of Nucleoside-5'-Triphosphates...- VI.2.2.3. Role of Protein Synthesis.- VI.3. Posttranscriptional Control.- VI.3.1. Synthesis and Supply of r-Proteins.- VI.3.2. The Role of Pre-rRNA Structure.- VI.3.3. The Role of 5 S rRNA.- VI.3.4. Critical Control Sites.- VI.3.4.1. Alternative Processing Pathways and Intranuclear Degradation of Preribosomes and Ribosomes.- VI.3.4.2. Release From the Nucleolus and Nucleo- Cytoplasmic Transport of Ribosomes.- VI.3.4.3. Turnover of Ribosomes.- VI.4. Autogeneous Regulation of Ribosome Biogenesis in Eukaryotes: A Model.- VI.5. Synopsis.- VII. Ribosome Biogenesis in the Life Cycle of Normal and Cancer Cells.- VII. 1. Nucleologenesis and Nucleololysis.- VII. 1.1. Nucleoli and Ribosome Biogenesis During the Mitotic Cycle.- VII. 1.2. Nucleologenesis.- VII. 1.3. Nuclyeololysis.- VII.2. Inhibition of Ribosome Biogenesis.- VII.2.1. Inhibitors Interacting With DNA and Chromatin.- VII.2.2. Inhibitors That Act on RNA Polymerases.- VII.2.3. Inhibitors of Nucleoside-5'-Triphosphate Formation.- VII.2.4. The Effects of Analogues Incorporated into Polyribonu- cleotide Chains.- VII.2.5. Inhibitors of Protein Synthesis.- VII.2.6. Interpretation of Nucleolar Alterations.- VII.2.6.1. Nucleolar Segregation.- VII.2.6.2. Nucleolar Spherical Bodies and Perichromatin Granules.- VII.2.6.3. Microspherules.- VII.2.6.4. Nucleolar Fragmentation.- VII.3. Growth Transitions.- VII.3.1. Modulation of Growth Rates in Yeasts.- VII.3.2. Activation of Lymphocytes.- VII.3.3. Growth Stimulation of Cultured Cells.- VII.3.4. Differentiation of Myoblasts in Culture.- VII.3.5. Regeneration of Rat Liver.- VII.4. Senescent and Cancer Cells.- VII.4.1. Senscent Cells and Tissues.- VII.4.2. Cancer Cells.- VII.5. Synopsis.- References.
514 citations
Cites background from "Sequence analysis of nuclear matrix..."
...An interesting observation showed that nuclear matrix DNA is about 5-fo1d enriched in rRNA genes (PARDOLL and VOGELSTEIN 1980)....
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Journal Article•
TL;DR: A nuclear matrix structure termed the nuclear matrix has been isolated and characterized in this article, which consists largely of protein with smaller amounts of RNA, DNA, carbohydrate, and phospholipid.
Abstract: A nuclear framework structure termed the nuclear matrix has been isolated and characterized. This matrix forms the major residual structure of isolated nuclei and consists largely of protein with smaller amounts of RNA, DNA, carbohydrate, and phospholipid. The nuclear matrix can be further resolved by combined treatment with DNase and RNase. The remaining nuclear protein structure, after extraction of 90 percent of the nuclear protein, 99.9 percent of the DNA, and 98 percent of the RNA and phospholipid, is termed the nuclear protein matrix. Electron microscopy of this final nuclear protein matrix reveals an interior framework structure composed of residual nucleolar structures associated with a granular and fibrous internal matrix structure. The internal matrix framework is derived from the interchromatinic structures of the nucleus, and is connected to a surrounding residual nuclear envelope layer containing residual nuclear pore complex structures. Sodium dodecyl sulfate-acrylamide gel electrophoresis of the nuclear matrix proteins demonstrates three major polypeptide fractions, P-1, P-2, and P-3, with average molecular weights of approximately 69,000, 66,000 and 62,000, as well as several minor polypeptides which migrate at approximately 50,000 and at higher molecular weights (>100,000). Polypeptides with molecular weights identical to those of P-1, P-2 and P-3 are also components of isolated nuclear envelopes and nucleoli, whereas isolated chromatin contains no detectable matrix polypeptides. This suggests that the major matrix polypeptides are localized in specific structural regions of the nucleus, i.e., nuclear envelope, nucleoli, and interchromatinic structures. The presence of cytochrome oxidase activity in the isolated nuclear matrix indicates that at least some integral proteins of the nuclear membrane are associated with the matrix.
508 citations
TL;DR: The observation that the transcriptionally active ovalbumin gene is preferentially associated with the nuclear matrix may have significant implications for gene expression and the organization of nuclear DNA into supercoiled-loop domains.
306 citations
TL;DR: Banks of ORI and MAR sequences are established and a large project of sequencing a large number of MARs is undertaken in an effort to determine classes of DNA sequences in these regulatory elements and to understand their role at the origins of replication and transcriptional enhancers.
Abstract: Polynuceosomes are constrained into loops or domains and are insulated from the effects of chromatin structure and torsional strain from flanking domains by the cross-complexation of matrix-attached regions (MARs) and matrix proteins. MARs or SARs have an average size of 500 bp, are spaced about every 30 kb, and are control elements maintaining independent realms of gene activity. A fraction of MARs may cohabit with core origins of replication (ORIs) and another fraction might cohabit with transcriptional enhancers. DNA replication, transcription, repair, splicing, and recombination seem to take place on the nuclear matrix. Classical AT-rich MARs have been proposed to anchor the core enhancers and core origins complexed with low abundancy transcription factors to the nuclear matrix via the cooperative binding to MARs of abundant classical matrix proteins (topoisomerase II, histone H1, lamins, SP120, ARBP, SATB1); this creates a unique nuclear microenvironment rich in regulatory proteins able to sustain transcription, replication, repair, and recombination. Theoretical searches and experimental data strongly support a model of activation of MARs and ORIs by transcription factors. A set of 21 characteristics are deduced or proposed for MAR/ORI sequences including their enrichment in inverted repeats, AT tracts, DNA unwinding elements, replication initiator protein sites, homooligonucleotide repeats (i.e., AAA, TTT, CCC), curved DNA, DNase l-hypersensitive sites, nucleosome-free stretches, polypurine stretches, and motifs with a potential for left-handed and triplex structures. We are establishing Banks of ORI and MAR sequences and have undertaken a large project of sequencing a large number of MARs in an effort to determine classes of DNA sequences in these regulatory elements and to understand their role at the origins of replication and transcriptional enhancers.
198 citations
TL;DR: It can be argued that theUnderstanding of eukaryotic rRNA processing is no less important than the understanding of mRNA maturation, since the capacity of a cell to carry out protein synthesis is controlled, in part, by the abundance of ribosomes.
Abstract: In summary, it can be argued that the understanding of eukaryotic rRNA processing is no less important than the understanding of mRNA maturation, since the capacity of a cell to carry out protein synthesis is controlled, in part, by the abundance of ribosomes. Processing of pre-rRNA is highly regulated, involving many cellular components acting either alone or as part of a complex. Some of these components are directly involved in the modification and cleavage of the precursor rRNA, while others direct the packaging of the rRNA into ribosome subunits. As is the case for pre-mRNA processing, snoRNPs are clearly involved in eukaryotic rRNA processing, and have been proposed to assemble with other proteins into at least one complex called a "processosome" (17), which carries out the ordered processing of the pre-rRNA and its assembly into ribosomes. The formation of a processing complex clearly makes possible the regulation required to coordinate the abundance of ribosomes with the physiological and developmental changes of a cell. It may be that eukaryotic rRNA processing is even more complex than pre-mRNA maturation, since pre-rRNA undergoes extensive nucleotide modification and is assembled into a complex structure called the ribosome. Undoubtedly, features of the eukaryotic rRNA-processing pathway have been conserved evolutionarily, and the genetic approach available in yeast research (6) should provide considerable knowledge that will be useful for other investigators working with higher eukaryotic systems. Interestingly, it was originally hoped that the extensive work and understanding of bacterial ribosome formation would provide a useful paradigm for the process in eukaryotes. However, although general features of ribosome structure and function are highly conserved between bacterial and eukaryotic systems, the basic strategy in ribosome biogenesis seems to be, for the most part, distinctly different. Thus, the detailed molecular mechanisms for rRNA processing in each kingdom will have to be independently deciphered in order to elucidate the features and regulation of this important process for cell survival.
196 citations
References
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TL;DR: A new method for isolation of high molecular weight DNA from eukaryotes is presented, which allows preparation of DNA from a variety of tissues and from cells which were more difficult to lyse until now.
Abstract: A new method for isolation of high molecular weight DNA from eukaryotes is presented. This procedure allows preparation of DNA from a variety of tissues such as calf thymus or human placenta and from cells which were more difficult to lyse until now (e.g. Crypthecodinium cuhnii, a dinoflagellate). The DNA obtained in such a way has an average molecular weight of about 200 X 10(6) d and contains very few, if any, single strand breaks.
2,524 citations
TL;DR: Direct support for the idea that regulation of gene activity underlies cell differentiation comes from evidence that much of the genome in higher cell types is inactive and that different ribonucleic acids are synthesized in different cell types.
Abstract: Cell differentiation is based almost certainly on the regulation of gene activity, so that for each state of differentiation a certain set of genes is active in transcription and other genes are inactive. The establishment of this concept (1) has depended on evidence
indicating that the cells of an organism generally contain identical genomes (2). Direct support for the idea that regulation of gene activity underlies cell differentiation
comes from evidence that much of the genome in higher cell
types is inactive (3) and that different ribonucleic acids (RNA) are synthesized in different cell types (4).
1,898 citations
TL;DR: The structural framework of the rat liver nucleus has been identified and consists of a nuclear protein matrix that is composed primarily of three acidic polypeptide fractions in the molecular weight range of 60–70,000 daltons.
983 citations
TL;DR: The thermal stabilities of RNA:DNA hybrids are substantially greater than those of DNA:DNA duplexes in aqueous electrolyte solutions containing high concentrations of formamide.
Abstract: The thermal stabilities of RNA:DNA hybrids are substantially greater than those of DNA:DNA duplexes in aqueous electrolyte solutions containing high concentrations of formamide. Association rates to form DNA:DNA duplexes and DNA:RNA hybrids have been measured in these solvents. There is a temperature range in which DNA:DNA rates are negligible and RNA:DNA rates close to optimal.
652 citations
TL;DR: Electron microscopic autoradiography shows that, as with intact nuclei, sites of DNA replication are distributed throughout the nuclear matrix, and a fixed site of DNA synthesis is proposed in which DNA replication complexes are anchored to thenuclear matrix.
624 citations