scholarly journals The Ndc80 complex targets Bod1 to human mitotic kinetochores

2017 ◽  
Author(s):  
Katharina Schleicher ◽  
Sara ten Have ◽  
Iain M. Porter ◽  
Jason R. Swedlow

AbstractRegulation of protein phosphatase activity by endogenous protein inhibitors is an important mechanism to control protein phosphorylation in cells. We recently identified Biorientation defective 1 (Bod1) as a small protein inhibitor of PP2A-B56. This phosphatase controls the amount of phosphorylation of mitotic kinetochore proteins and thus the establishment of load-bearing chromosome-spindle attachments in time for accurate separation of sister chromatids in mitosis. The PP2A-B56 regulatory protein Bod1 directly localises to mitotic kinetochores and is required for correct segregation of mitotic chromosomes. In this report, we have probed the spatio-temporal regulation of Bod1 during mitotic progression. Kinetochore localisation of Bod1 increases from nuclear envelope breakdown until metaphase. Phosphorylation of Bod1 at threonine 95 (T95), which increases Bod1’s binding and inhibition of PP2A-B56, peaks in prometaphase, when PP2A-B56 localisation to kinetochores is highest. Kinetochore targeting of Bod1 depends on the outer kinetochore protein Ndc80 and not PP2A-B56. Additionally, Bod1 depletion leads to a defect in the phosphorylation kinetics of Ndc80 at serine 55 (S55) within its N-terminus. Therefore, Ndc80 recruits a phosphatase inhibitor to kinetochores which directly feeds forward to regulate Ndc80 phosphorylation and the stability of the attachments of microtubules to kinetochores.

Open Biology ◽  
2017 ◽  
Vol 7 (11) ◽  
pp. 170099 ◽  
Author(s):  
Katharina Schleicher ◽  
Michael Porter ◽  
Sara ten Have ◽  
Ramasubramanian Sundaramoorthy ◽  
Iain M. Porter ◽  
...  

Regulation of protein phosphatase activity by endogenous protein inhibitors is an important mechanism to control protein phosphorylation in cells. We recently identified Biorientation defective 1 (Bod1) as a small protein inhibitor of protein phosphatase 2A containing the B56 regulatory subunit (PP2A-B56). This phosphatase controls the amount of phosphorylation of several kinetochore proteins and thus the establishment of load-bearing chromosome-spindle attachments in time for accurate separation of sister chromatids in mitosis. Like PP2A-B56, Bod1 directly localizes to mitotic kinetochores and is required for correct segregation of mitotic chromosomes. In this report, we have probed the spatio-temporal regulation of Bod1 during mitotic progression. Kinetochore localization of Bod1 increases from nuclear envelope breakdown until metaphase. Phosphorylation of Bod1 at threonine 95 (T95), which increases Bod1's binding to and inhibition of PP2A-B56, peaks in prometaphase when PP2A-B56 localization to kinetochores is highest. We demonstrate here that kinetochore targeting of Bod1 depends on the outer kinetochore protein Ndc80 and not PP2A-B56. Crucially, Bod1 depletion functionally affects Ndc80 phosphorylation at the N-terminal serine 55 (S55), as well as a number of other phosphorylation sites within the outer kinetochore, including Knl1 at serine 24 and 60 (S24, S60), and threonine T943 and T1155 (T943, T1155). Therefore, Ndc80 recruits a phosphatase inhibitor to kinetochores which directly feeds forward to regulate Ndc80, and Knl1 phosphorylation, including sites that mediate the attachment of microtubules to kinetochores.


1977 ◽  
Vol 16 (04) ◽  
pp. 157-162 ◽  
Author(s):  
C. Schümichen ◽  
B. Mackenbrock ◽  
G. Hoffmann

SummaryThe bone-seeking 99mTc-Sn-pyrophosphate compound (compound A) was diluted both in vitro and in vivo and proved to be unstable both in vitro and in vivo. However, stability was much better in vivo than in vitro and thus the in vitro stability of compound A after dilution in various mediums could be followed up by a consecutive evaluation of the in vivo distribution in the rat. After dilution in neutral normal saline compound A is metastable and after a short half-life it is transformed into the other 99mTc-Sn-pyrophosphate compound A is metastable and after a short half-life in bone but in the kidneys. After dilution in normal saline of low pH and in buffering solutions the stability of compound A is increased. In human plasma compound A is relatively stable but not in plasma water. When compound B is formed in a buffering solution, uptake in the kidneys and excretion in urine is lowered and blood concentration increased.It is assumed that the association of protons to compound A will increase its stability at low concentrations while that to compound B will lead to a strong protein bond in plasma. It is concluded that compound A will not be stable in vivo because of a lack of stability in the extravascular space, and that the protein bond in plasma will be a measure of its in vivo stability.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuping Li ◽  
Xiaoju Liang ◽  
Xuguo Zhou ◽  
Yu An ◽  
Ming Li ◽  
...  

AbstractGlycyrrhiza, a genus of perennial medicinal herbs, has been traditionally used to treat human diseases, including respiratory disorders. Functional analysis of genes involved in the synthesis, accumulation, and degradation of bioactive compounds in these medicinal plants requires accurate measurement of their expression profiles. Reverse transcription quantitative real-time PCR (RT-qPCR) is a primary tool, which requires stably expressed reference genes to serve as the internal references to normalize the target gene expression. In this study, the stability of 14 candidate reference genes from the two congeneric species G. uralensis and G. inflata, including ACT, CAC, CYP, DNAJ, DREB, EF1, RAN, TIF1, TUB, UBC2, ABCC2, COPS3, CS, R3HDM2, were evaluated across different tissues and throughout various developmental stages. More importantly, we investigated the impact of interactions between tissue and developmental stage on the performance of candidate reference genes. Four algorithms, including geNorm, NormFinder, BestKeeper, and Delta Ct, were used to analyze the expression stability and RefFinder, a comprehensive software, provided the final recommendation. Based on previous research and our preliminary data, we hypothesized that internal references for spatio-temporal gene expression are different from the reference genes suited for individual factors. In G. uralensis, the top three most stable reference genes across different tissues were R3HDM2, CAC and TUB, while CAC, CYP and ABCC2 were most suited for different developmental stages. CAC is the only candidate recommended for both biotic factors, which is reflected in the stability ranking for the spatio (tissue)-temporal (developmental stage) interactions (CAC, R3HDM2 and DNAJ). Similarly, in G. inflata, COPS3, R3HDM2 and DREB were selected for tissues, while RAN, COPS3 and CS were recommended for developmental stages. For the tissue-developmental stage interactions, COPS3, DREB and ABCC2 were the most suited reference genes. In both species, only one of the top three candidates was shared between the individual factors and their interactions, specifically, CAC in G. uralensis and COPS3 in G. inflata, which supports our overarching hypothesis. In summary, spatio-temporal selection of reference genes not only lays the foundation for functional genomics research in Glycyrrhiza, but also facilitates these traditional medicinal herbs to reach/maximize their pharmaceutical potential.


2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Veniamin Zheleznov ◽  
Aleksey Golikov ◽  
Tatiana Sokolnitskaya ◽  
Sergey Ivannikov

Abstract The sorption kinetics of uranyl ions micro-quantities from fluoride solutions by nanostructured materials with anatase mesoporous structures has been studied. Using the model of competitive sorption of ions and positively charged complexes of uranyl ion on deprotonated hydroxyl groups of an anatase, kinetic curves of changes in the ratio of ionic forms of uranium in solution were calculated. Modeling was carried out under the assumption of a two-stage mechanism of uranium complex ions sorption. The modeling considered the influence of the uranyl ion carbonate complexes formation. The shift in equilibrium among ionic forms of uranyl correlates with the stability of the complexes in solution.


2020 ◽  
Vol 48 (12) ◽  
pp. 6583-6596
Author(s):  
Akiko Fujimura ◽  
Yuki Hayashi ◽  
Kazashi Kato ◽  
Yuichiro Kogure ◽  
Mutsuro Kameyama ◽  
...  

Abstract The nucleolus is a membrane-less nuclear structure that disassembles when cells undergo mitosis. During mitosis, nucleolar factors are thus released from the nucleolus and dynamically change their subcellular localization; however, their functions remain largely uncharacterised. Here, we found that a nucleolar factor called nucleolar protein 11 (NOL11) forms a protein complex with two tryptophan-aspartic acid (WD) repeat proteins named WD-repeat protein 43 (WDR43) and Cirhin in mitotic cells. This complex, referred to here as the NWC (NOL11-WDR43-Cirhin) complex, exists in nucleoli during interphase and translocates to the periphery of mitotic chromosomes, i.e., perichromosomal regions. During mitotic progression, both the congression of chromosomes to the metaphase plate and sister chromatid cohesion are impaired in the absence of the NWC complex, as it is required for the centromeric enrichment of Aurora B and the associating phosphorylation of histone H3 at threonine 3. These results reveal the characteristics of a novel protein complex consisting of nucleolar proteins, which is required for regulating kinetochores and centromeres to ensure faithful chromosome segregation.


2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Gui-chen Li ◽  
Chong-chong Qi ◽  
Yuan-tian Sun ◽  
Xiao-lin Tang ◽  
Bao-quan Hou

The kinetics of fluid-solid coupling during immersion is an important topic of investigation in rock engineering. Two rock types, sandstone and mudstone, are selected in this work to study the correlation between the softening characteristics of the rocks and moisture content. This is achieved through detailed studies using scanning electron microscopy, shear tests, and evaluation of rock index properties during exposure to different moisture contents. An underground roadway excavation is simulated by dynamic finite element modeling to analyze the effect of moisture content on the stability of the roadway. The results show that moisture content has a significant effect on shear properties reduction of both sandstone and mudstone, which must thus be considered in mining or excavation processes. Specifically, it is found that the number, area, and diameter of micropores, as well as surface porosity, increase with increasing moisture content. Additionally, stress concentration is negatively correlated with moisture content, while the influenced area and vertical displacement are positively correlated with moisture content. These findings may provide useful input for the design of underground roadways.


MRS Bulletin ◽  
1990 ◽  
Vol 15 (1) ◽  
pp. 41-47 ◽  
Author(s):  
Nikola Kallay

The adhesion of particles at solid surfaces in liquid media has attracted the attention of scientists because of its various applications as well as the theoretical significance of the processes involved. Early studies were characterized either by poorly defined systems or limited by the properties of a few morphologically well-defined model colloids, such as latex dispersions. Consequently, results were either of semiquantitative nature or were related to some specific cases, which eluded general conclusions. New methods for preparing uniform particles of different compositions, shapes, and sizes make it possible to approach the problem in a more comprehensive manner. For example, to demonstrate difficulties caused by polydispersity, it is sufficient to mention that the electrostatic interaction energy between a plane surface and a particle is approximately proportional to the particle radius, yet the rate of deposition depends exponentially on the height of the energy barrier.In principle, static and dynamic approaches may be employed in the study of particle adhesion. The static method yields the force required to detach an adhered particle, while kinetic investigations of attachment and detachment give the rates of the respective processes. Both methods offer information on the stability of the system in terms of the bond strength of adhered solids. For small colloid particles, which are the subject of thermal random Brownian motion, the dynamic approach is more appropriate. This article emphasizes the kinetics of deposition and detachment of small colloid particles in liquid media.


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