scholarly journals Partially Processed pre-rRNA Is Preserved in Association with Processing Components in Nucleolus-derived Foci during Mitosis

1998 ◽  
Vol 9 (9) ◽  
pp. 2407-2422 ◽  
Author(s):  
Miroslav Dundr ◽  
Mark O.J. Olson
Keyword(s):  
Molecular Weight ◽  
Internal Transcribed Spacer ◽  
Rnase P ◽  
Rrna Processing ◽  
Rnase Mrp ◽  
External Transcribed Spacer ◽  
Early Anaphase ◽  
Late Telophase ◽  
Processing Site

Previous studies showed that components implicated in pre-rRNA processing, including U3 small nucleolar (sno)RNA, fibrillarin, nucleolin, and proteins B23 and p52, accumulate in perichromosomal regions and in numerous mitotic cytoplasmic particles, termed nucleolus-derived foci (NDF) between early anaphase and late telophase. The latter structures were analyzed for the presence of pre-rRNA by fluorescence in situ hybridization using probes for segments of pre-rRNA with known half-lives. The NDF did not contain the short-lived 5′-external transcribed spacer (ETS) leader segment upstream from the primary processing site in 47S pre-rRNA. However, the NDF contained sequences from the 5′-ETS core, 18S, internal transcribed spacer 1 (ITS1), and 28S segments and also had detectable, but significantly reduced, levels of the 3′-ETS sequence. Northern analyses showed that in mitotic cells, the latter sequences were present predominantly in 45S-46S pre-rRNAs, indicating that high-molecular weight processing intermediates are preserved during mitosis. Two additional essential processing components were also found in the NDF: U8 snoRNA and hPop1 (a protein component of RNase MRP and RNase P). Thus, the NDF appear to be large complexes containing partially processed pre-rRNA associated with processing components in which processing has been significantly suppressed. The NDF may facilitate coordinated assembly of postmitotic nucleoli.

scholarly journals Basic Domains Target Protein Subunits of the RNase MRP Complex to the Nucleolus Independently of Complex Association

2001 ◽  
Vol 12 (11) ◽  
pp. 3680-3689 ◽  
Author(s):  
Hans van Eenennaam ◽  
Annemarie van der Heijden ◽  
Rolf J. R. J. Janssen ◽  
Walther J. van Venrooij ◽  
Ger J. M. Pruijn
Keyword(s):  
Protein Composition ◽  
Rnase P ◽  
Nucleolar Localization ◽  
Rrna Processing ◽  
Protein Subunits ◽  
Trna Processing ◽  
Rnase Mrp ◽  
Human Rnase ◽  
The Difference ◽  
Active Transport Process

The RNase MRP and RNase P ribonucleoprotein particles both function as endoribonucleases, have a similar RNA component, and share several protein subunits. RNase MRP has been implicated in pre-rRNA processing and mitochondrial DNA replication, whereas RNase P functions in pre-tRNA processing. Both RNase MRP and RNase P accumulate in the nucleolus of eukaryotic cells. In this report we show that for three protein subunits of the RNase MRP complex (hPop1, hPop4, and Rpp38) basic domains are responsible for their nucleolar accumulation and that they are able to accumulate in the nucleolus independently of their association with the RNase MRP and RNase P complexes. We also show that certain mutants of hPop4 accumulate in the Cajal bodies, suggesting that hPop4 traverses through these bodies to the nucleolus. Furthermore, we characterized a deletion mutant of Rpp38 that preferentially associates with the RNase MRP complex, giving a first clue about the difference in protein composition of the human RNase MRP and RNase P complexes. On the basis of all available data on nucleolar localization sequences, we hypothesize that nucleolar accumulation of proteins containing basic domains proceeds by diffusion and retention rather than by an active transport process. The existence of nucleolar localization sequences is discussed.

The organization of ribosomal RNA processing correlates with the distribution of nucleolar snRNAs

1996 ◽  
Vol 109 (6) ◽  
pp. 1241-1251 ◽  
Author(s):  
A.F. Beven ◽  
R. Lee ◽  
M. Razaz ◽  
D.J. Leader ◽  
J.W. Brown ◽  
...  
Keyword(s):  
Small Bodies ◽  
Transcript Processing ◽  
Coiled Bodies ◽  
External Transcribed Spacer ◽  
Immunofluorescence Labelling ◽  
Late Telophase ◽  
Ribosomal Rna Processing ◽  
Fibrillar Component ◽  
High Degree

We have analyzed the organization of pre-rRNA processing by confocal microscopy in pea root cell nucleoli using a variety of probes for fluorescence in situ hybridization and immunofluorescence. Our results show that transcript processing within the nucleolus is spatially highly organized. Probes to the 5′ external transcribed spacer (ETS) and first internal transcribed spacer (ITS1) showed that the excision of the ETS occurred in a sub-region of the dense fibrillar component (DFC), whereas the excision of ITS1 occurred in the surrounding region, broadly corresponding to the granular component. In situ labelling with probes to the snoRNAs U3 and U14, and immunofluorescence labelling with antibodies to fibrillarin and SSB1 showed a high degree of coincidence with the ETS pattern, confirming that ETS cleavage and 18 S rRNA production occur in the DFC. ETS, U14, fibrillarin and SSB1 showed a fine substructure within the DFC comprising closely packed small foci, whereas U3 appeared more diffuse throughout the DFC. A third snoRNA, 7–2/MRP, was localised to the region surrounding the ETS, in agreement with its suggested role in ITS1 cleavage. All three snoRNAs were also frequently observed in numerous small foci in the nucleolar vacuoles, but none was detectable in coiled bodies. Antibodies to fibrillarin and SSB1 labelled coiled bodies strongly, though neither protein was detected in the nucleolar vacuoles. During mitosis, all the components analyzed, including pre-rRNA, were dispersed through the cell at metaphase, then became concentrated around the periphery of all the chromosomes at anaphase, before being localized to the developing nucleoli at late telophase. Pre-rRNA (ETS and ITS1 probes), U3 and U14 were also concentrated into small bodies, presumed to be pre-nucleolar bodies at anaphase.

scholarly journals Hierarchical recruitment of ribosomal proteins and assembly factors remodels nucleolar pre-60S ribosomes

2018 ◽  
Vol 217 (7) ◽  
pp. 2503-2518 ◽  
Author(s):  
Stephanie Biedka ◽  
Jelena Micic ◽  
Daniel Wilson ◽  
Hailey Brown ◽  
Luke Diorio-Toth ◽  
...  
Keyword(s):  
Internal Transcribed Spacer ◽  
Ribosomal Proteins ◽  
Ribosome Biogenesis ◽  
Large Subunit ◽  
Ribosome Assembly ◽  
Rrna Processing ◽  
Endonucleolytic Cleavage ◽  
Cryo Electron Microscopy ◽  
External Transcribed Spacer ◽  
Assembly Factors

Ribosome biogenesis involves numerous preribosomal RNA (pre-rRNA) processing events to remove internal and external transcribed spacer sequences, ultimately yielding three mature rRNAs. Removal of the internal transcribed spacer 2 spacer RNA is the final step in large subunit pre-rRNA processing and begins with endonucleolytic cleavage at the C2 site of 27SB pre-rRNA. C2 cleavage requires the hierarchical recruitment of 11 ribosomal proteins and 14 ribosome assembly factors. However, the function of these proteins in C2 cleavage remained unclear. In this study, we have performed a detailed analysis of the effects of depleting proteins required for C2 cleavage and interpreted these results using cryo–electron microscopy structures of assembling 60S subunits. This work revealed that these proteins are required for remodeling of several neighborhoods, including two major functional centers of the 60S subunit, suggesting that these remodeling events form a checkpoint leading to C2 cleavage. Interestingly, when C2 cleavage is directly blocked by depleting or inactivating the C2 endonuclease, assembly progresses through all other subsequent steps.

Monitoring Polymer-Assisted Mechanochemical Cocrystallisation Through in Situ X-Ray Powder Diffraction

2020 ◽  
Author(s):  
Luzia S. Germann ◽  
Sebastian T. Emmerling ◽  
Manuel Wilke ◽  
Robert E. Dinnebier ◽  
Mariarosa Moneghini ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Powder Diffraction ◽  
Reaction Rate ◽  
Polymer Additives ◽  
Time Resolved ◽  
X Ray

Time-resolved mechanochemical cocrystallisation studies have so-far focused solely on neat and liquid-assisted grinding. Here, we report the monitoring of polymer-assisted grinding reactions using <i>in situ</i> X-ray powder diffraction, revealing that reaction rate is almost double compared to neat grinding and independent of the molecular weight and amount of used polymer additives.<br>

scholarly journals Achieving Good Protection on Ultra-High Molecular Weight Polythene by In Situ Growth of Amorphous Carbon Film

Coatings ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 584
Author(s):  
Rui Dang ◽  
Liqiu Ma ◽  
Shengguo Zhou ◽  
Deng Pan ◽  
Bin Xia
Keyword(s):  
Molecular Weight ◽  
Epitaxial Growth ◽  
High Molecular Weight ◽  
Amorphous Carbon ◽  
Transition Layer ◽  
In Situ Growth ◽  
Good Protection ◽  
Ultra High Molecular Weight ◽  
Carbon Plasma

Ultra-high molecular weight polythene (UHMWPE), with outstanding characteristics, is widely applied in modern industry, while it is also severely limited by its inherent shortcomings, which include low hardness, poor wear resistance, and easy wear. Implementation of feasible protection on ultra-high molecular weight polythene to overcome its shortcomings would be of significance. In the present study, amorphous carbon (a-C) film was fabricated on ultra-high molecular weight polythene (UHMWPE) to provide good protection, and the relevant growth mechanism of a-C film was revealed by controlling carbon plasma currents. The results showed the in situ transition layer, in the form of chemical bonds, was formed between the UHMWPE substrate and the a-C film with the introduction of carbon plasma, which provided strong adhesion, and then the a-C film continued epitaxial growth on the in situ transition layer with the treatment of carbon plasma. This in situ growth of a-C film, including the in situ transition layer and the epitaxial growth layer, significantly improved the wetting properties, mechanical properties, and tribological properties of UHMWPE. In particular, good protection by in situ growth a-C film on UHMWPE was achieved during sliding wear.

Wear behavior of in situ polymerized carbon nanotube/ultra high molecular weight polyethylene composites

2013 ◽  
Vol 21 (9) ◽  
pp. 965-970 ◽  
Author(s):  
Hong-Jo Park ◽  
Jihun Kim ◽  
Yongsok Seo ◽  
Junho Shim ◽  
Moon-Yong Sung ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Carbon Nanotube ◽  
High Molecular Weight ◽  
Wear Behavior ◽  
Ultra High Molecular Weight ◽  
Polyethylene Composites

scholarly journals Preparation of Porous Polymers by ”in Situ Precipitation” Using Low Molecular Weight Gelators

2002 ◽  
Vol 34 (6) ◽  
pp. 474-477 ◽  
Author(s):  
Masahiro Suzuki ◽  
Yasuhiko Sakakibara ◽  
Satoshi Kobayashi ◽  
Mutsumi Kimura ◽  
Hirofusa Shirai ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Porous Polymers ◽  
Low Molecular Weight ◽  
Low Molecular Weight Gelators

Preparation and Properties of the Polystyrene/Alumina Nanocomposites

2012 ◽  
Vol 557-559 ◽  
pp. 519-522
Author(s):  
Xu Man Wang ◽  
Cai Ning Zhang
Keyword(s):  
Molecular Weight ◽  
Thermal Resistance ◽  
Coupling Agent ◽  
Silane Coupling Agent ◽  
In Situ Polymerization ◽  
Average Molecular Weight ◽  
Experimental Results ◽  
Solvent Resistance ◽  
Silane Coupling

Silane coupling agent KH-570 was applied to modify the surface capability of the alumina (Al2O3). The modified Al2O3were dispersed in styrene. The in-situ polymerization was used to prepare the polystyrene/alumina (PS/Al2O3) composites, in which azodiisobutyronitrile (AIBN) was used as initiator. FTIR, DSC and TG were applied to characterize the prepared composites. The solvent resistance, thermal resistance of the composites and the average molecular weight of PS in PS/Al2O3nanocomposites were studied. The experimental results demonstrated that the solvent resistance of PS/Al2O3nanocomposites was improved by the adding of Al2O3nanoparticles. The thermal resistance of the composites increased with the increasing of the Al2O3content. Meanwhile, the molecular weight of PS in the composites increased with the increasing of the Al2O3content.

Research on In Situ Anionic Polymerization of PA6/PI Alloy

2011 ◽  
Vol 295-297 ◽  
pp. 21-25
Author(s):  
Hong Kai Zhao ◽  
Li Guang Xiao ◽  
Hong Jie Wang
Keyword(s):  
Molecular Weight ◽  
Bisphenol A ◽  
High Performance ◽  
Hot Spot ◽  
Microscopic Analysis ◽  
Infrared Analysis ◽  
Elongation At Break ◽  
System Temperature ◽  
Lamellar Crystals

High performance trend of plastics has become a hot spot of current research. Select bisphenol A dianhydride and bisphenol A diamine with excellent water resistance as the reactant monomers to obtain anhydride-terminated polyimide with very high molecular weight by two-step polymerization, graft the active radicals of acyl caprolactam using the activity of anhydride and obtain PI modified nylon resin by polymerization.When the system temperature is above 160 °C and the added modifiers are greater than 10%, the system viscosity increases very fast; when the system temperature reaches 140 °C and the added modifiers are at 5%, the system viscosity increases very slowly. It is proved that the reaction in each above step is successful through infrared analysis. The mechanical properties of modified PI nylon increases with the increase of consumption and molecular weight of polyimide, when the molecular weight is selected to be about 8000~10000 and the adding amount is 10wt%~15 wt%, the tensile strength reaches over 85MPa, the notch impact strength is increased to 19.6kJ.m-2 and the elongation at break reaches 18%, which are remarkably better than general engineering plastics.Through microscopic analysis, the molecules of polyimide does not enter crystallization phase of nylon resin, but forms compact lamellar crystals existing in nylon matrix.

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