Showing papers on "Nucleolar chromatin published in 1976"

The nucleolar structure.

Abstract: Publisher Summary The nucleolus is organized at the nucleolus-organizing region of the chromosomes, which are generally visible as secondary constriction regions in metaphase chromosomes. The chromatin within the constriction region is lost at interphase inside the nucleolar mass. The chromatin is highly extended at this stage. At the ultrastructural level, the nucleolus has at least three components: (1) a granular component consisting mainly of ribonucleoproteins (RNP) granules—pars granulosa, (2) a fibrillar component, consisting of RNP fibrils—pars fibrosa, and (3) chromatin elements. Chromatin elements may be present in three forms: (1) nucleolus-associated chromatin, which most likely does not take part in nucleolus formation; however, the possibility of its association with condensed inactive ribosomal cistrons, at least in some cells, cannot be overruled at present, (2) septalike intranucleolar chromatin, and (3) isolated or dispersed intranucleolar chromatin threads. Intranucleolar chromatin is often associated with the pars fibrosa. Identical components can also be found in isolated nucleoli. Studies on the nucleoli in giant chromosomes indicate that the intranucleolar chromatin in these nucleoli is present as puffs of different sizes. The nucleolar chromatin is not an autonomous structure of the nucleolus but is a continuous structure and part of the nucleolar chromosome.

Regulation of transcription of genes of ribosomal rna during amphibian oogenesis. A biochemical and morphological study.

Abstract: Natural changes in the transcription of rRNA genes were studied in nucleoli from three oogenic stages of the newt Triturus alpestris with electron microscope, auto-radiographic, and biochemical techniques From determinations of the uridine triphosphate pool sizes and [3H]uridine uptake, phosphorylation, and incorporation into 28S and 18S rRNAs in vivo it was estimated that the rate of rRNA synthesis was about 001% in previtellogenic oocytes and 13% in mature oocytes when compared to midvitellogenesis Spread preparations of nucleoli showed significant morphological changes in the transcriptional complexes The total number of lateral fibrils, ie, ribonucleoproteins containing the nascent rRNA precursor, were drastically decreased in stages of reduced synthetic activity This indicates that rRNA synthesis is regulated primarily at the level of transcription The resulting patterns of fibril coverage of the nucleolar chromatin axes revealed a marked heterogeneity On the same nucleolar axis occurred matrix units that were completely devoid of lateral fibrils, matrix units that were almost fully covered with lateral fibrils, and various forms of matrix units with a range of lateral fibril densities intermediate between the two extremes Granular particles that were tentatively identified as RNA polymerase molecules were not restricted to the transciptional complexes They were observed, although less regularly and separated by greater distances, in untranscribed spacer regions as well as in untranscribed gene intercepts The results show that the pattern of transcriptional control of rRNA genes differs widely in different genes, even in the same genetic unit

Homogeneity and heterogeneity of sizes of transcriptional units and spacer regions in nucleolar genes of acetabularia

Abstract: The arrangement of genes of precursor molecules for ribosomal RNA (pre-rRNA) in primary nuclei from two green algae species, Acetabularia mediterranea and A. major, has been analyzed in an electron microscope study. The pattern of transcriptional units in individual strands of nucleolar chromatin was investigated using spread and posi­ tively stained preparations. The rDNA pattern is not uniform but differs in different strands. The predominant type of nucleolar chromatin exhibits a high degree of homogeneity in the sequence of matrix units (intercepts covered with fibrilst hat contain the pre-rRNA) and fibril-free spacer intercepts. Substantial differences, however, are observed between the patterns in different strands. In addition, there is evidence in some strands for intraaxial heterogeneity of both spacer and matrix units. The following major types can be distinguished: type la, ca. 2 micrometer long matrix units, extremely short spacer intercepts in A. mediterranea (ca. 1 micrometer long ones in A. major), completely homogeneous distribution; type Ib, as type la but with intercalated, isolated, significantly shorter and/or longer matrix units; type lIa, matrix unit sizes as in type la, but much longer spacer intercepts, high degree of homogeneity; type Ill, largely heterogeneous arrangements of matrix and spacer units of varying sizes. The matrix unit data are compared with the sizes of pre-rRNA as determined by polyacrylamide gelelectrophoresis under denaturing and non-denaturing conditions. The findings are discussed in relation to recent observations in amphibia and insects and with respect to current concepts of the species-specificity of rDNA arrangements.

Circular dichroic studies of the DNA and RNA of nucleoli.

Abstract: Circular dichroism (CD) in the 240-300-nm region was used to study the conformation of DNA and RNA complexed with proteins in isolated nucleoli form HeLa cells. Deoxyribonuclease or ribonuclease digestion was employed to obtain (1) the individual CD spectra of nucleolar DNA or RNA in complex form with proteins, or in free form; and (2) the experimental CD baseline correction to exclude contributions from nonnucleic acid sources such as light scattering artifacts and proteins. The CD spectrum of nucleolar DNA in DNA-protein complexes was highly reduced in ellipticity in comparison with protein-free DNA. It showed a positive peak at 283 nm with a molar ellipticity [theta]283 = 1200 deg cm2 dmol-1 and a crossover at 262 nm. Addition of sodium dodecylsulfate shifted the peak to 276 nm with [theta]276 8000 deg cm2 dmol-1 and a crossover at 254 nm. The CD spectrum of nucleolar RNA in RNA-protein complexes was also reduced in comparison with protein-free RNA, showing a peak at 269 nm ([theta]269 = 6900 deg cm2 dmol-1), and a crossover at 250 nm. Addition of sodium dodecyl sulfate shifted the peak to 265 nm with [theta]265 = 18 000 deg cm2 dmol-1 and a crossover at 246 nm. The low ellipticity of both nucleolar DNA and RNA when complexed with proteins was increased by treatment with sodium chloride, urea, or heparin. This suggests that some ionic, hydrophobic, and hydrogen bondings are involved in the nucleic acid-protein interaction in nucleolar chromatin similar to that observed in nuclear chromatin.

Ultrastructural aspects of nucleolar fibrillar centres in meristematic cells ofAllium cepa

Abstract: This paper deals with the fine structure of the fibrillar centres of the nucleolus inAllium cepa cells in ultrathin, sections of in vivo fixed roots. The ultrastructural observations have allowed us to consider each nucleolar fibrillar centre as an active zone in the nucleolar chromatin loop, and to propose a possible model for the organization of the different components of the nucleolus within it.

Homochromatographic and immunological analysis of controls of nucleolar gene function.

Abstract: To compare regulation of nucleolar function of tumors and other tissues, it was necessary to develop assays of the fidelity of ribosomal DNA readouts. For this purpose, homochromatography analyses of complete T1 ribonuclease digestion products of the in vivo labeled 45 S preribosomal RNA were compared with those of 18S and of 28 S ribosomal RNA. Homochromatography analysis of the in vitro readout product of isolated nucleoli showed the presence of many large marker nucleotides of the in vivo 45 S preribosomal RNA. Moreover, no other large oligonucleotides were detected. The in vitro readout product of nucleolar chromatin had the same T1 ribonuclease digestion products, including the large marker of oligonucleotides. However, the in vitro readout product of nucleolar DNA contained no large marker T1 ribonuclease oligonucleotides. These results indicate that the fidelity of nucleolar readouts is controlled by regulatory proteins of the nucleolar chromatin. Differences were found in nucleolar proteins of normal rat liver and Novikoff hepatoma by immunological analyses. The possibility exists that differences in readout rates of tumor and other nucleoli are related to the protein difference detected by these immunological studies.