Nucleolus

About: Nucleolus is a research topic. Over the lifetime, 5873 publications have been published within this topic receiving 232435 citations. The topic is also known as: GO:0005730 & cell nucleolus.

Subnuclear localization of the active variant surface glycoprotein gene expression site in Trypanosoma brucei (antigenic variationyin situ hybridizationynucleolusyRNA polymerase I)

Abstract: In Trypanosoma brucei, transcription by RNA polymerase II and 5* capping of messenger RNA are uncou- pled: a capped spliced leader is trans spliced to every RNA. This decoupling makes it possible to have protein-coding gene transcription driven by RNA polymerase I. Indeed, indirect evidence suggests that the genes for the major surface glyco- proteins, variant surface glycoproteins (VSGs) in blood- stream-form trypanosomes, are transcribed by RNA polymer- ase I. In a single trypanosome, only one VSG expression site is maximally transcribed at any one time, and it has been speculated that transcription takes place at a unique site within the nucleus, perhaps in the nucleolus. We tested this by using f luorescence in situ hybridization. With probes that cover about 50 kb of the active 221 expression site, we detected nuclear transcripts of this site in a single f luorescent spot, which did not colocalize with the nucleolus. Analysis of marker gene-tagged active expression site DNA by f luorescent DNA in situ hybridization confirmed the absence of associa- tion with the nucleolus. Even an active expression site in which the promoter had been replaced by an rDNA promoter did not colocalize with the nulceolus. As expected, marker genes inserted in the rDNA array predominantly colocalize with the nucleolus, whereas the tubulin gene arrays do not. We con- clude that transcription of the active VSG expression site does not take place in the nucleolus.

Presence of Pre-rRNAs before Activation of Polymerase I Transcription in the Building Process of Nucleoli during Early Development of Xenopus laevis

Abstract: During the early development of Xenopus laevis, we followed in individual nuclei the formation of a nucleolus by examining simultaneously its structural organization and its transcriptional competence. Three distinct situations were encountered with different frequencies during development. During the first period of general transcriptional quiescence, the transcription factor UBF of maternal origin, was present in most nuclei at the ribosomal gene loci. In contrast, fibrillarin, a major protein of the processing machinery, was found in multiple prenucleolar bodies (PNBs) whereas nucleolin was dispersed largely in the nucleoplasm. During the second period, for most nuclei these PNBs had fused into two domains where nucleolin concentrated, generating a structure with most features expected from a transcriptionally competent nucleolus. However, RNA polymerase I–dependent transcription was not detected using run-on in situ assays whereas unprocessed ribosomal RNAs were observed. These RNAs were found to derive from a maternal pool. Later, during a third period, an increasing fraction of the nuclei presented RNA polymerase I–dependent transcription. Thus, the structural organization of the nucleolus preceded its transcriptional competence. We conclude that during the early development of X. laevis, the organization of a defined nucleolar structure, is not associated with the transcription process per se but rather with the presence of unprocessed ribosomal RNAs.

The material continuity and individuality of the somatic chromosomes of Ambystoma tigrinum, with special reference to the nucleolus as a chromosomal component

Abstract: Two features of this study have furnished direct evidence for the material continuity and individuality of the chromosomes in the epithelial cells of the larvae of Ambystoma tigrinum (1) the tracing of the complete history of a specific chromosome, the nucleolus-containing chromosome, throughout the successive stages of mitosis and interkinesis, (2) the following of the exact parallelism in behavior (during mitosis) of the chromosomes in a given nuclear group with that in a single chromosome which has been isolated in the cytoplasm from the main chromosomal complex. Each of the two nucleoli is subterminally located in the short arm of one of two particular V-shaped chromosomes. The latter are probably homologues. 78: In this subterminal chromosomal position the nucleolus exhibits in fixed and stained preparations an achromatic character in late prophase, metaphase, anaphase and early telophase, while it presents itself as a chromatic element in middle telophase, late telophase, interkinesis and early prophase. The nucleolus is not a simple homogeneous body, but rather a complex structure, the nature of which is revealed in the study of its evolution during the chromatic phase of its cycle in mitosis.

Isolation and proteomic characterization of the major proteins of the nucleolin-binding ribonucleoprotein complexes.

Abstract: Nucleolin (NCL) is one of the most abundant nucleolar proteins of exponentially growing eukaryotic cells. It is known to interact only transiently with rRNA and preribosomal particles and not to be detectable in mature cytoplasmic ribosomes, and is believed to function as multi-protein complexes during ribosome biogenesis and maturation. However, those multiprotein complexes remain only partially characterized due to the difficulty of conventional protein analysis methods. Here we report isolation of NCL-binding protein complex and its proteomic characterization with the use of an analytical method based on matrix-assisted laser desorption/ionization-time of flight analysis coupled with searching peptide mass databases. The NCL-binding protein complex was isolated by immunoprecipitation with anti-Flag antibody from human kidney 293 cells that were transfected with the Flag-tagged NCL gene, and showed RNA integrity for holding their protein constituents. Interaction between NCL and its binding complex was disrupted by an RNA oligonucleotide with a NCL recognition element, indicating that NCL binds to the ribonucleoprotein (RNP) complex mainly through the sequence specific protein-RNA interaction. We confirmed that an RNA-binding domain of NCL alone was sufficient to hold the entire NCL-binding RNP complex, indicating the strict binding specificity of NCL to the isolated RNP complex in 293 cells. We identified forty ribosomal proteins from both the large and small subunits, and twenty nonribosomal proteins. These results together suggest that the isolated NCL-binding RNP complex is a preribosomal particle present in the nucleolus of 293 cells.