When rDNA transcription is arrested during mitosis, UBF is still associated with non-condensed rDNA

1997 ◽  
Vol 110 (19) ◽  
pp. 2429-2440 ◽  
Author(s):  
J. Gebrane-Younes ◽  
N. Fomproix ◽  
D. Hernandez-Verdun
Keyword(s):  
Secondary Constriction ◽  
Three Dimensional ◽  
Ribosomal Gene ◽  
Rdna Transcription ◽  
Transcription Machinery ◽  
Late Anaphase ◽  
Early Anaphase ◽  
Dimensional Distribution ◽  
Open Question ◽  
Kinetics Of

The mechanisms that control inactivation of ribosomal gene (rDNA) transcription during mitosis is still an open question. To investigate this fundamental question, the precise timing of mitotic arrest was established. In PtK1 cells, rDNA transcription was still active in prophase, stopped in prometaphase until early anaphase, and activated in late anaphase. Because rDNA transcription can still occur in prophase and late anaphase chromosomes, the kinetics of rDNA condensation during mitosis was questioned. The conformation of the rDNA was analyzed by electron microscopy from the G2/M transition to late anaphase in the secondary constriction, the chromosome regions where the rDNAs are clustered. Whether at transcribing or non-transcribing stages, non-condensed rDNA was observed in addition to axial condensed rDNA. Thus, the persistence of this non-condensed rDNA during inactive transcription argues in favor of the fact that mitotic inactivation is not the consequence of rDNA condensation. Analysis of the three-dimensional distribution of the rDNA transcription factor, UBF, revealed that it was similar at each stage of mitosis in the secondary constriction. In addition, the colocalization of UBF with non-condensed rDNA was demonstrated. This is the first visual evidence of the association of UBF with non-condensed rDNA. As we previously reported that the rDNA transcription machinery remained assembled during mitosis, the colocalization of rDNA fibers with UBF argues in favor of the association of the transcription machinery with certain rDNA copies even in the absence of transcription. If this hypothesis is correct, it can be assumed that condensation of rDNA as well as dissociation of the transcription machinery from rDNA cannot explain the arrest of rDNA transcription during mitosis. It is proposed that modifications of the transcription machinery occurring in prometaphase could explain the arrest of transcription, while reverse modifications in late anaphase could explain activation.

The mitotically phosphorylated form of the transcription termination factor TTF-1 is associated with the repressed rDNA transcription machinery

1999 ◽  
Vol 112 (19) ◽  
pp. 3259-3268 ◽  
Author(s):  
V. Sirri ◽  
P. Roussel ◽  
D. Hernandez-Verdun
Keyword(s):  
Transcription Termination ◽  
Nucleolar Organizer ◽  
Ribosomal Gene ◽  
Nucleolar Organizer Regions ◽  
Rdna Transcription ◽  
Mitotic Chromosomes ◽  
Termination Factor ◽  
Transcription Machinery ◽  
Phosphorylated Form ◽  
Transcription Termination Factor

The transcription termination factor TTF-1 exerts two functions in ribosomal gene (rDNA) transcription: facilitating initiation and mediating termination of transcription. Using HeLa cells, we show that TTF-1 protein is colocalized with the active transcription machinery in the nucleolus and also with the inactive machinery present in certain mitotic nucleolar organizer regions (NORs) when rDNA transcription is repressed. We also show that TTF-1 is specifically phosphorylated during mitosis in a manner dependent on the cdc2-cyclin B kinase pathway and on an okadaic acid-sensitive phosphatase. Interestingly, the mitotically phosphorylated form of TTF-1 appearing at the G(2)/M transition phase was more easily solubilized than was the interphase form. This indicates that the chromatin-binding affinity of TTF-1 appears to be different in mitotic chromosomes compared to the interphase nucleolus. Correlated with this, the other DNA-binding factor, UBF, which interferes with chromatin conformation in the rDNA promoter, was more strongly bound to rDNA during mitosis than at interphase. The reorganization of the mitotic rDNA promoter might be induced by phosphorylation of certain components of the rDNA transcription machinery and participate in silencing of rDNA during mitosis.

Cytoplasmic Nucleoloids of Lilium Male Reproductive Cells Contain rDNA Transcripts and Share Features of Development with Nucleoli

1991 ◽  
Vol 100 (1) ◽  
pp. 109-118 ◽  
Author(s):  
S. SATO ◽  
K. JONES ◽  
J. DE LOS DIOS ALCHE ◽  
H. G. DICKINSON
Keyword(s):  
In Situ Hybridisation ◽  
Ribosomal Gene ◽  
Gene Probe ◽  
Developmental Pathway ◽  
Rdna Transcription ◽  
Rna Sequences ◽  
Tetrad Stage ◽  
Late Anaphase ◽  
Identical Fashion

Nucleolus-like inclusions in the meiocytes of Lilium sp. have been investigated at the light and electronmicroscope levels, with respect both to their composition and their formation during the meiotic divisions. Those present in the cytoplasm (cytoplasmic nucleoloids) first appear as small structures late in anaphase I. Coincidentally, small nuclear nucleolus like inclusions (NLBs) develop on the surface of the chromosomes, and sometimes in the region between chromosomes as they are carried into the daughter nuclei. The cytoplasmic nucleoloids increase in size from late anaphase I to the dyad stage, but disappear before metaphase II. They redevelop late in anaphase II and persist, increasing in size, until the tetrad stage. The NLBs also increase in size over this period. Ultrastructural investigation indicates that both the nascent nucleoloids and NLBs are fibrous in texture as they arise in late anaphase. Elsewhere in the cytoplasm, however, large accumulations of amorphous electron-opaque material also aggregate. This material becomes associated with the nucleoloids and the NLBs during the period of their enlargement. Nucleoloids late in development, and post-telophase nucleoli are not invested by these amorphous masses. Interestingly, cytoplasmic nucleoloids and NLBs react in an identical fashion to a range of cytochemical treatments and, using in situ hybridisation at the electron-microscope level, they have been demonstrated to contain RNA sequences homologous with a wheat ribosomal gene probe. Cytoplasmic nucleoloids are thus interpreted as following an identical developmental pathway to nucleoli except in that, perhaps owing to high levels of rDNA transcription during meiosis, they condense in the cytoplasm rather than the nucleoplasm.

scholarly journals In Vivo Release of Mitotic Silencing of Ribosomal Gene Transcription Does Not Give Rise to Precursor Ribosomal RNA Processing

2000 ◽  
Vol 148 (2) ◽  
pp. 259-270 ◽  
Author(s):  
Valentina Sirri ◽  
Pascal Roussel ◽  
Danièle Hernandez-Verdun
Keyword(s):  
Ribosomal Rna ◽  
Kinase Activity ◽  
Ribosomal Gene ◽  
Cyclin B ◽  
Rrna Synthesis ◽  
Rrna Processing ◽  
Rdna Transcription ◽  
Transcription Machinery ◽  
Mitotic Cells

Nuclear RNA transcription is repressed when eukaryotic cells enter mitosis. Here, we found that the derepression of ribosomal gene (rDNA) transcription that normally takes place in telophase may be induced in prometaphase, metaphase, and anaphase mitotic HeLa cells, and therefore appears not to be dependent on completion of mitosis. We demonstrate for the first time that in vivo inhibition of the cdc2– cyclin B kinase activity is sufficient to give rise to okadaic acid–sensitive dephosphorylation of the mitotically phosphorylated forms of components of the rDNA transcription machinery, and consequently to restore rDNA transcription in mitotic cells. These results, showing that during mitosis the rDNA transcription machinery is maintained repressed by the cdc2–cyclin B kinase activity, provide an in vivo demonstration of the cell cycle–dependent regulation of rDNA transcription. Interestingly in mitotic cells, the newly synthesized 47S precursor ribosomal RNA (pre-rRNA) is not processed into the mature rRNAs, indicating that rDNA transcription and pre-rRNA processing may be uncoupled. Moreover this suggests that inhibition of the cdc2– cyclin B kinase is not sufficient to activate the 47S pre-rRNA processing machinery and/or to induce its relocalization at the level of newly synthesized 47S pre-rRNA. This in vivo approach provides new possibilities to investigate the correlation between pre-rRNA synthesis and pre-rRNA processing when the nucleolus reforms.

Three-dimensional distribution of ribulose-1, 5-bisphosphate carboxylase/oxygemase during the early development of proplastids in dark-grown Euglena cells transferred to an inorganic medium

1990 ◽  
Vol 48 (3) ◽  
pp. 906-907
Author(s):  
Tomoko Ehara ◽  
Shuji Sumida ◽  
Tetsuaki Osafune ◽  
Eiji Hase
Keyword(s):  
Cell Suspension ◽  
Euglena Gracilis ◽  
Carbon Materials ◽  
Three Dimensional ◽  
Peripheral Region ◽  
Plastid Development ◽  
Bisphosphate Carboxylase ◽  
Inorganic Medium ◽  
Ribulose 1 ◽  
Dimensional Distribution

As shown previously, Euglena cells grown in Hutner’s medium in the dark without agitation accumulate wax as well as paramylum, and contain proplastids showing no internal structure except for a single prothylakoid existing close to the envelope. When the cells are transferred to an inorganic medium containing ammonium salt and the cell suspension is aerated in the dark, the wax was oxidatively metabolized, providing carbon materials and energy 23 for some dark processes of plastid development. Under these conditions, pyrenoid-like structures (called “pro-pyrenoids”) are formed at the sites adjacent to the prolamel larbodies (PLB) localized in the peripheral region of the proplastid. The single prothylakoid becomes paired with a newly formed prothylakoid, and a part of the paired prothylakoids is extended, with foldings, in to the “propyrenoid”. In this study, we observed a concentration of RuBisCO in the “propyrenoid” of Euglena gracilis strain Z using immunoelectron microscopy.

scholarly journals A Novel Pseudomonas geniculata AGE Family Epimerase/Isomerase and Its Application in d-Mannose Synthesis

Foods ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1809
Author(s):  
Zhanzhi Liu ◽  
Ying Li ◽  
Jing Wu ◽  
Sheng Chen
Keyword(s):  
3D Structure ◽  
Three Dimensional ◽  
Optimal Temperature ◽  
Reaction Conditions ◽  
Enzymatic Production ◽  
Physiological Properties ◽  
Potential Applications ◽  
Kinetics Of ◽  
Optimal Reaction ◽  
Gene Age

d-mannose has exhibited excellent physiological properties in the food, pharmaceutical, and feed industries. Therefore, emerging attention has been applied to enzymatic production of d-mannose due to its advantage over chemical synthesis. The gene age of N-acetyl-d-glucosamine 2-epimerase family epimerase/isomerase (AGEase) derived from Pseudomonas geniculata was amplified, and the recombinant P. geniculata AGEase was characterized. The optimal temperature and pH of P. geniculata AGEase were 60 °C and 7.5, respectively. The Km, kcat, and kcat/Km of P. geniculata AGEase for d-mannose were 49.2 ± 8.5 mM, 476.3 ± 4.0 s−1, and 9.7 ± 0.5 s−1·mM−1, respectively. The recombinant P. geniculata AGEase was classified into the YihS enzyme subfamily in the AGE enzyme family by analyzing its substrate specificity and active center of the three-dimensional (3D) structure. Further studies on the kinetics of different substrates showed that the P. geniculata AGEase belongs to the d-mannose isomerase of the YihS enzyme. The P. geniculata AGEase catalyzed the synthesis of d-mannose with d-fructose as a substrate, and the conversion rate was as high as 39.3% with the d-mannose yield of 78.6 g·L−1 under optimal reaction conditions of 200 g·L−1d-fructose and 2.5 U·mL−1P. geniculata AGEase. This novel P. geniculata AGEase has potential applications in the industrial production of d-mannose.

scholarly journals Dip angles of active faults from the surface to the seismogenic zone inferred from a 2D numerical analysis of visco-elasto-plastic models: a case study for the Osaka Plain

2021 ◽  
Vol 73 (1) ◽  
Author(s):  
Hayami Nishiwaki ◽  
Takamoto Okudaira ◽  
Kazuhiko Ishii ◽  
Muneki Mitamura
Keyword(s):  
Urban Areas ◽  
Active Faults ◽  
Three Dimensional ◽  
Seismogenic Zone ◽  
Earthquake Ground Motion ◽  
Convex Curve ◽  
Seismic Reflection Profile ◽  
Geological Structures ◽  
Dimensional Distribution ◽  
Dip Angle

AbstractThe geometries (i.e., dip angles) of active faults from the surface to the seismogenic zone are the most important factors used to evaluate earthquake ground motion, which is crucial for seismic hazard assessments in urban areas. In Osaka, a metropolitan city in Japan, there are several active faults (e.g., the Uemachi and Ikoma faults), which are inferred from the topography, the attitude of active faults in surface trenches, the seismic reflection profile at shallow depths (less than 2 km), and the three-dimensional distribution of the Quaternary sedimentary layers. The Uemachi and Ikoma faults are N–S-striking fault systems with total lengths of 42 km and 38 km, respectively, with the former being located ~ 12 km west of the latter; however, the geometries of each of the active faults within the seismogenic zone are not clear. In this study, to examine the geometries of the Uemachi and Ikoma faults from the surface to the seismogenic zone, we analyze the development of the geological structures of sedimentary layers based on numerical simulations of a two-dimensional visco-elasto-plastic body under a horizontal compressive stress field, including preexisting high-strained weak zones (i.e., faults) and surface sedimentation processes, and evaluate the relationship between the observed geological structures of the Quaternary sediments (i.e., the Osaka Group) in the Osaka Plain and the model results. As a result, we propose geometries of the Uemachi and Ikoma faults from the surface to the seismogenic zone. When the friction coefficient of the faults is ~ 0.5, the dip angles of the Uemachi and Ikoma faults near the surface are ~ 30°–40° and the Uemachi fault has a downward convex curve at the bottom of the seismogenic zone, but does not converge to the Ikoma fault. Based on the analysis in this study, the dip angle of the Uemachi fault zone is estimated to be approximately 30°–40°, which is lower than that estimated in the previous studies. If the active fault has a low angle, the width of the fault plane is long, and thus the estimated seismic moment will be large.

scholarly journals Simulation and measurement of air quality in the traffic congestion area

2021 ◽  
Vol 31 (1) ◽  
Author(s):  
Shin Yu ◽  
Chang Tang Chang ◽  
Chih Ming Ma
Keyword(s):  
Air Quality ◽  
Traffic Congestion ◽  
Traffic Management ◽  
Reference Model ◽  
Dispersion Model ◽  
Fluid Model ◽  
Three Dimensional ◽  
Vehicle Speed ◽  
Management Plans ◽  
Dimensional Distribution

AbstractThe traffic congestion in the Hsuehshan tunnel and at the Toucheng interchange has led to traffic-related air pollution with increasing concern. To ensure the authenticity of our simulation, the concentration of the last 150 m in Hsuehshan tunnel was simulated using the computational fluid dynamics fluid model. The air quality at the Toucheng interchange along a 2 km length highway was simulated using the California Line Source Dispersion Model. The differences in air quality between rush hours and normal traffic conditions were also investigated. An unmanned aerial vehicle (UAV) with installed PM2.5 sensors was developed to obtain the three-dimensional distribution of pollutants. On different roads, during the weekend, the concentrations of pollutants such as SOx, CO, NO, and PM2.5 were observed to be in the range of 0.003–0.008, 7.5–15, 1.5–2.5 ppm, and 40–80 μg m− 3, respectively. On weekdays, the vehicle speed and the natural wind were 60 km h− 1 and 2.0 m s− 1, respectively. On weekdays, the SOx, CO, NO, and PM2.5 concentrations were found to be in the range of 0.002–0.003, 3–9, 0.7–1.8 ppm, and 35–50 μg m− 3, respectively. The UAV was used to verify that the PM2.5 concentrations of vertical changes at heights of 9.0, 7.0, 5.0, and 3.0 m were 45–48, 30–35, 25–30, and 50–52 μg m− 3, respectively. In addition, the predicted PM2.5 concentrations were 40–45, 25–30, 45–48, and 45–50 μg m− 3 on weekdays. These results provide a reference model for environmental impact assessments of long tunnels and traffic jam-prone areas. These models and data are useful for transportation planners in the context of creating traffic management plans.

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