scholarly journals Chromosome segregation fidelity is controlled by small changes in phospho-occupancy at the kinetochore-microtubule interface

2021 ◽  
Author(s):  
Thomas J. Kucharski ◽  
Rufus Hards ◽  
Kristina M. Godek ◽  
Scott A. Gerber ◽  
Duane A. Compton
Keyword(s):  
Error Correction ◽  
Chromosome Segregation ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Cyclin B1 ◽  
Quantitative Mass Spectrometry ◽  
Kinetochore Protein ◽  
Microtubule Attachment ◽  
Different Types ◽  
Narrow Dynamic Range

SummaryKinetochore protein phosphorylation promotes the correction of erroneous microtubule attachments to ensure faithful chromosome segregation during cell division. Determining how phosphorylation executes error correction requires an understanding of whether kinetochore substrates are completely (i.e. all-or-none) or only fractionally phosphorylated. Using quantitative mass spectrometry (MS), we measured phospho-occupancy on the conserved kinetochore protein Hec1 (NDC80) that directly binds microtubules. None of the positions measured exceeded ∼50% phospho-occupancy, and the cumulative phospho-occupancy changed by only ∼20% in response to changes in microtubule attachment status. The narrow dynamic range of phospho-occupancy is maintained by ongoing phosphatase activity. Further, both Cdk1-Cyclin B1 and Aurora kinases phosphorylate Hec1 to enhance error correction in response to different types of microtubule attachment errors. Thus, networks of kinases and phosphatases maintain low inherent phospho-occupancy to promote microtubule attachment to kinetochores while providing for high sensitivity of kinetochore-microtubule attachments to very small changes in phospho-occupancy to ensure high mitotic fidelity.

scholarly journals Delineating the contribution of Spc105-bound PP1 to spindle checkpoint silencing and kinetochore microtubule attachment regulation

2019 ◽  
Vol 218 (12) ◽  
pp. 3926-3942 ◽  
Author(s):  
Babhrubahan Roy ◽  
Vikash Verma ◽  
Janice Sim ◽  
Adrienne Fontan ◽  
Ajit P. Joglekar
Keyword(s):  
Cell Division ◽  
Error Correction ◽  
Chromosome Segregation ◽  
Protein Phosphatase ◽  
Spindle Assembly Checkpoint ◽  
Spindle Checkpoint ◽  
Spindle Assembly ◽  
Protein Phosphatase 1 ◽  
Microtubule Attachment ◽  
Error Correction Mechanism

Accurate chromosome segregation during cell division requires the spindle assembly checkpoint (SAC), which detects unattached kinetochores, and an error correction mechanism that destabilizes incorrect kinetochore–microtubule attachments. While the SAC and error correction are both regulated by protein phosphatase 1 (PP1), which silences the SAC and stabilizes kinetochore–microtubule attachments, how these distinct PP1 functions are coordinated remains unclear. Here, we investigate the contribution of PP1, docked on its conserved kinetochore receptor Spc105/Knl1, to SAC silencing and attachment regulation. We find that Spc105-bound PP1 is critical for SAC silencing but dispensable for error correction; in fact, reduced PP1 docking on Spc105 improved chromosome segregation and viability of mutant/stressed states. We additionally show that artificially recruiting PP1 to Spc105/Knl1 before, but not after, chromosome biorientation interfered with error correction. These observations lead us to propose that recruitment of PP1 to Spc105/Knl1 is carefully regulated to ensure that chromosome biorientation precedes SAC silencing, thereby ensuring accurate chromosome segregation.

scholarly journals Aurora B switches relative strength between kinetochore–microtubule attachment modes to promote error correction

2018 ◽  
Author(s):  
Harinath Doodhi ◽  
Taciana Kasciukovic ◽  
Lesley Clayton ◽  
Tomoyuki U. Tanaka
Keyword(s):  
Error Correction ◽  
Chromosome Segregation ◽  
Relative Strength ◽  
Lateral Side ◽  
Aurora B ◽  
Spindle Poles ◽  
Microtubule Attachment ◽  
Attachment Strength ◽  
Dam1 Complex

AbstractFor proper chromosome segregation, sister kinetochores must interact with microtubules from opposite spindle poles; this is called bi-orientation. To establish bi-orientation prior to chromosome segregation, any aberrant kinetochore–microtubule interaction must be resolved (error correction) by Aurora B kinase that phosphorylates outer kinetochore components. Aurora B differentially regulates kinetochore attachment to the microtubule plus end and its lateral side (end-on and lateral attachment, respectively). However, it is still not fully understood how kinetochore–microtubule interactions are exchanged during error correction. Here we reconstituted the kinetochore–microtubule interface of budding yeast in vitro by attaching the Ndc80 complexes (Ndc80C) to nanobeads. These Ndc80C–nanobeads recapitulated in vitro the lateral and end-on attachments of authentic kinetochores, on dynamic microtubules loaded with the Dam1 complex. This in vitro assay enabled the direct comparison of lateral and end-on attachment strength and showed that Dam1 phosphorylation by Aurora B makes the end-on attachment weaker than the lateral attachment. We suggest that the Dam1 phosphorylation weakens interaction with the Ndc80 complex, disrupts the end-on attachment and promotes the exchange to a new lateral attachment, leading to error correction. Our study reveals a fundamental mechanism of error correction for establishment of bi-orientation.

scholarly journals Kinetic Variability at The 5’ UTR of Bacterial mRNA Leads to Narrow Dynamic Range and High Sensitivity in 30S Ribosome Binding

2020 ◽  
Vol 118 (3) ◽  
pp. 546a
Author(s):  
Shiba S. Dandpat ◽  
Sujay Ray ◽  
Surajit Chatterjee ◽  
Nils G. Walter
Keyword(s):  
Dynamic Range ◽  
High Sensitivity ◽  
Ribosome Binding ◽  
Narrow Dynamic Range

scholarly journals Reconstitution reveals two paths of force transmission through the kinetochore

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Grace E Hamilton ◽  
Luke A Helgeson ◽  
Cameron L Noland ◽  
Charles L Asbury ◽  
Yoana N Dimitrova ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Error Correction ◽  
Chromosome Segregation ◽  
Force Transmission ◽  
Daughter Cells ◽  
Self Assembling ◽  
Microtubule Attachment ◽  
Gain And Loss ◽  
Dynamic Microtubule

Partitioning duplicated chromosomes equally between daughter cells is a microtubule-mediated process essential to eukaryotic life. A multi-protein machine, the kinetochore, drives chromosome segregation by coupling the chromosomes to dynamic microtubule tips, even as the tips grow and shrink through the gain and loss of subunits. The kinetochore must harness, transmit, and sense mitotic forces, as a lack of tension signals incorrect chromosome-microtubule attachment and precipitates error correction mechanisms. But though the field has arrived at a ‘parts list’ of dozens of kinetochore proteins organized into subcomplexes, the path of force transmission through these components has remained unclear. Here we report reconstitution of functional Saccharomyces cerevisiae kinetochore assemblies from recombinantly expressed proteins. The reconstituted kinetochores are capable of self-assembling in vitro, coupling centromeric nucleosomes to dynamic microtubules, and withstanding mitotically relevant forces. They reveal two distinct pathways of force transmission and Ndc80c recruitment.

Backscattered Electron Imaging of GaAs/AlGaAs Super Lattice Structures with an Ultra-High- Resolution SEM

1988 ◽  
Vol 46 ◽  
pp. 204-205
Author(s):  
Kazumichi Ogura ◽  
Michael M. Kersker
Keyword(s):  
High Resolution ◽  
Layer Structure ◽  
High Sensitivity ◽  
Beam Current ◽  
Electron Beam Current ◽  
Lattice Structures ◽  
Super Lattice ◽  
Different Types ◽  
Backscattered Electron ◽  
Electron Imaging

Backscattered electron (BE) images of GaAs/AlGaAs super lattice structures were observed with an ultra high resolution (UHR) SEM JSM-890 with an ultra high sensitivity BE detector. Three different types of super lattice structures of GaAs/AlGaAs were examined. Each GaAs/AlGaAs wafer was cleaved by a razor after it was heated for approximately 1 minute and its crosssectional plane was observed.First, a multi-layer structure of GaAs (100nm)/AlGaAs (lOOnm) where A1 content was successively changed from 0.4 to 0.03 was observed. Figures 1 (a) and (b) are BE images taken at an accelerating voltage of 15kV with an electron beam current of 20pA. Figure 1 (c) is a sketch of this multi-layer structure corresponding to the BE images. The various layers are clearly observed. The differences in A1 content between A1 0.35 Ga 0.65 As, A1 0.4 Ga 0.6 As, and A1 0.31 Ga 0.69 As were clearly observed in the contrast of the BE image.

Slow-Scan CCD Observation of Backscattering Patterns in SEM

1992 ◽  
Vol 50 (2) ◽  
pp. 1308-1309
Author(s):  
F. Ouyang ◽  
D. A. Ray ◽  
O. L. Krivanek
Keyword(s):  
Electron Beam ◽  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Dynamic Range ◽  
High Sensitivity ◽  
Lens System ◽  
Wide Dynamic Range ◽  
Peltier Cooler ◽  
Diffraction Patterns ◽  
Scanning Electron

Electron backscattering Kikuchi diffraction patterns (BKDP) reveal useful information about the structure and orientation of crystals under study. With the well focused electron beam in a scanning electron microscope (SEM), one can use BKDP as a microanalysis tool. BKDPs have been recorded in SEMs using a phosphor screen coupled to an intensified TV camera through a lens system, and by photographic negatives. With the development of fiber-optically coupled slow scan CCD (SSC) cameras for electron beam imaging, one can take advantage of their high sensitivity and wide dynamic range for observing BKDP in SEM.We have used the Gatan 690 SSC camera to observe backscattering patterns in a JEOL JSM-840A SEM. The CCD sensor has an active area of 13.25 mm × 8.83 mm and 576 × 384 pixels. The camera head, which consists of a single crystal YAG scintillator fiber optically coupled to the CCD chip, is located inside the SEM specimen chamber. The whole camera head is cooled to about -30°C by a Peltier cooler, which permits long integration times (up to 100 seconds).

Development of imaging plate for a TEM and its characteristics

1989 ◽  
Vol 47 ◽  
pp. 100-101
Author(s):  
N. Mori ◽  
T. Oikawa ◽  
Y. Harada ◽  
J. Miyahara ◽  
T. Matsuo
Keyword(s):  
Dynamic Range ◽  
Protective Layer ◽  
High Sensitivity ◽  
Imaging Plate ◽  
Phosphor Layer ◽  
Imaging Device ◽  
X Ray Imaging ◽  
New Type ◽  
Basic Characteristics ◽  
Reading System

The Imaging Plate (IP) is a new type imaging device, which was developed for diagnostic x ray imaging. We have reported that usage of the IP for a TEM has many merits; those are high sensitivity, wide dynamic range, and good linearity. However in the previous report the reading system was prototype drum-type-scanner, and IP was also experimentally made, which phosphor layer was 50μm thick with no protective layer. So special care was needed to handle them, and they were used only to make sure the basic characteristics. In this article we report the result of newly developed reading, printing system and high resolution IP for practical use. We mainly discuss the characteristics of the IP here. (Precise performance concerned with the reader and other system are reported in the other article.)Fig.1 shows the schematic cross section of the IP. The IP consists of three parts; protective layer, phosphor layer and support.

scholarly journals High-frequency rectifiers based on type-II Dirac fermions

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Libo Zhang ◽  
Zhiqingzi Chen ◽  
Kaixuan Zhang ◽  
Lin Wang ◽  
Huang Xu ◽  
...  
Keyword(s):  
High Frequency ◽  
Dynamic Range ◽  
High Sensitivity ◽  
High Dynamic Range ◽  
Anisotropy Ratio ◽  
Dirac Fermions ◽  
Topological States ◽  
Topological Semimetals ◽  
Polarized Surface ◽  
Symmetry Protected

AbstractThe advent of topological semimetals enables the exploitation of symmetry-protected topological phenomena and quantized transport. Here, we present homogeneous rectifiers, converting high-frequency electromagnetic energy into direct current, based on low-energy Dirac fermions of topological semimetal-NiTe2, with state-of-the-art efficiency already in the first implementation. Explicitly, these devices display room-temperature photosensitivity as high as 251 mA W−1 at 0.3 THz in an unbiased mode, with a photocurrent anisotropy ratio of 22, originating from the interplay between the spin-polarized surface and bulk states. Device performances in terms of broadband operation, high dynamic range, as well as their high sensitivity, validate the immense potential and unique advantages associated to the control of nonequilibrium gapless topological states via built-in electric field, electromagnetic polarization and symmetry breaking in topological semimetals. These findings pave the way for the exploitation of topological phase of matter for high-frequency operations in polarization-sensitive sensing, communications and imaging.

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