scholarly journals Chromosomal attachments set length and microtubule number in the Saccharomyces cerevisiae mitotic spindle

2014 ◽  
Vol 25 (25) ◽  
pp. 4034-4048 ◽  
Author(s):  
Natalie J. Nannas ◽  
Eileen T. O’Toole ◽  
Mark Winey ◽  
Andrew W. Murray
Keyword(s):  
Electron Microscopy ◽  
Saccharomyces Cerevisiae ◽  
Simple Model ◽  
Mitotic Spindle ◽  
Cell Types ◽  
Yeast Saccharomyces Cerevisiae ◽  
Spindle Poles ◽  
Length Regulation ◽  
Spindle Microtubules ◽  
Different Cell Types

The length of the mitotic spindle varies among different cell types. A simple model for spindle length regulation requires balancing two forces: pulling, due to micro­tubules that attach to the chromosomes at their kinetochores, and pushing, due to interactions between microtubules that emanate from opposite spindle poles. In the budding yeast Saccharomyces cerevisiae, we show that spindle length scales with kinetochore number, increasing when kinetochores are inactivated and shortening on addition of synthetic or natural kinetochores, showing that kinetochore–microtubule interactions generate an inward force to balance forces that elongate the spindle. Electron microscopy shows that manipulating kinetochore number alters the number of spindle microtubules: adding extra kinetochores increases the number of spindle microtubules, suggesting kinetochore-based regulation of microtubule number.

scholarly journals Kinetochores and chromatin diminution in early embryos of Parascaris univalens.

1992 ◽  
Vol 118 (1) ◽  
pp. 23-32 ◽  
Author(s):  
C Goday ◽  
J M González-García ◽  
M R Esteban ◽  
G Giovinazzo ◽  
S Pimpinelli
Keyword(s):  
Mitotic Spindle ◽  
Cell Types ◽  
Entire Length ◽  
Mitotic Chromosomes ◽  
Chromatin Diminution ◽  
Kinetic Activity ◽  
Early Embryos ◽  
Spindle Microtubules ◽  
Egg Shell ◽  
Different Cell Types

In Parascaris the mitotic chromosomes of gonial germline cells are holocentric and possess a continuous kinetochore along their entire length. By contrast, in meiotic cells, the centromeric activity is restricted to the heterochromatic tips where direct insertion of spindle microtubules into chromatin without any kinetochore plate is seen. In the presomatic cells of early embryos, which undergo heterochromatin elimination, only euchromatin shows kinetic activity. After developing a technique to separate the very resistant egg shell from the embryos, we studied the cell divisions during early embryogenesis by immunochemical and EM approaches. The results reported here show that in presomatic cells microtubules bind only the euchromatin where a continuous kinetochore plate is present. We also report observations suggesting that the binding of the long kinetochores to the mitotic spindle initiates to a limited number of sites and extends along the entire length, during chromosome condensation. The existence of different centromere stages in different cell types, rends Parascaris chromosomes a very good model to study centromere organization.

Centrosomes and mitotic spindle poles: a recent liaison?

2015 ◽  
Vol 43 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Pavithra L. Chavali ◽  
Isabel Peset ◽  
Fanni Gergely
Keyword(s):  
Cell Division ◽  
Mitotic Spindle ◽  
Potential Benefit ◽  
Cell Types ◽  
Daughter Cells ◽  
Spindle Poles ◽  
Pericentriolar Material ◽  
Mitotic Spindle Assembly ◽  
Spindle Microtubules ◽  
Ordered Assembly

Centrosomes comprise two cylindrical centrioles embedded in the pericentriolar material (PCM). The PCM is an ordered assembly of large scaffolding molecules, providing an interaction platform for proteins involved in signalling, trafficking and most importantly microtubule nucleation and organization. In mitotic cells, centrosomes are located at the spindle poles, sites where spindle microtubules converge. However, certain cell types and organisms lack centrosomes, yet contain focused spindle poles, highlighting that despite their juxtaposition in cells, centrosomes and mitotic spindle poles are distinct physical entities. In the present paper, we discuss the origin of centrosomes and summarize their contribution to mitotic spindle assembly and cell division. We then describe the key molecular players that mediate centrosome attachment to mitotic spindle poles and explore why co-segregation of centrosomes and spindle poles into daughter cells is of potential benefit to organisms.

Study of the Molecular Architecture of Keratin Filaments by Electron Microscopy

1982 ◽  
Vol 40 ◽  
pp. 118-119
Author(s):  
U. Aebi ◽  
P. Rew ◽  
T.-T. Sun
Keyword(s):  
Electron Microscopy ◽  
Structural Organization ◽  
Cell Types ◽  
Structural Features ◽  
Molecular Architecture ◽  
The Past ◽  
Keratin Filaments ◽  
Different Types ◽  
10 Nm Filaments ◽  
Different Cell Types

Various types of intermediate-sized (10-nm) filaments have been found and described in many different cell types during the past few years. Despite the differences in the chemical composition among the different types of filaments, they all yield common structural features: they are usually up to several microns long and have a diameter of 7 to 10 nm; there is evidence that they are made of several 2 to 3.5 nm wide protofilaments which are helically wound around each other; the secondary structure of the polypeptides constituting the filaments is rich in ∞-helix. However a detailed description of their structural organization is lacking to date.

scholarly journals Stu1p Is Physically Associated with β-Tubulin and Is Required for Structural Integrity of the Mitotic Spindle

2002 ◽  
Vol 13 (6) ◽  
pp. 1881-1892 ◽  
Author(s):  
Hongwei Yin ◽  
Liru You ◽  
Danielle Pasqualone ◽  
Kristen M. Kopski ◽  
Tim C. Huffaker
Keyword(s):  
Mitotic Spindle ◽  
Structural Integrity ◽  
Spindle Pole ◽  
Temperature Sensitive ◽  
Yeast Saccharomyces Cerevisiae ◽  
Spindle Poles ◽  
Balance Of Forces ◽  
Related Proteins ◽  
Β Tubulin

Formation of the bipolar mitotic spindle relies on a balance of forces acting on the spindle poles. The primary outward force is generated by the kinesin-related proteins of the BimC family that cross-link antiparallel interpolar microtubules and slide them past each other. Here, we provide evidence that Stu1p is also required for the production of this outward force in the yeast Saccharomyces cerevisiae. In the temperature-sensitive stu1–5mutant, spindle pole separation is inhibited, and preanaphase spindles collapse, with their previously separated poles being drawn together. The temperature sensitivity of stu1–5 can be suppressed by doubling the dosage of Cin8p, a yeast BimC kinesin–related protein. Stu1p was observed to be a component of the mitotic spindle localizing to the midregion of anaphase spindles. It also binds to microtubules in vitro, and we have examined the nature of this interaction. We show that Stu1p interacts specifically with β-tubulin and identify the domains required for this interaction on both Stu1p and β-tubulin. Taken together, these findings suggest that Stu1p binds to interpolar microtubules of the mitotic spindle and plays an essential role in their ability to provide an outward force on the spindle poles.

scholarly journals Role of POT1 in Human Cancer

Cancers ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 2739 ◽  
Author(s):  
Yangxiu Wu ◽  
Rebecca C. Poulos ◽  
Roger R. Reddel
Keyword(s):  
Human Cancer ◽  
Cell Types ◽  
Cancer Therapies ◽  
Telomeric Dna ◽  
Targeted Cancer Therapies ◽  
Length Regulation ◽  
Tumor Types ◽  
Different Cell Types ◽  
Essential Subunit

Telomere abnormalities facilitate cancer development by contributing to genomic instability and cellular immortalization. The Protection of Telomeres 1 (POT1) protein is an essential subunit of the shelterin telomere binding complex. It directly binds to single-stranded telomeric DNA, protecting chromosomal ends from an inappropriate DNA damage response, and plays a role in telomere length regulation. Alterations of POT1 have been detected in a range of cancers. Here, we review the biological functions of POT1, the prevalence of POT1 germline and somatic mutations across cancer predisposition syndromes and tumor types, and the dysregulation of POT1 expression in cancers. We propose a framework for understanding how POT1 abnormalities may contribute to oncogenesis in different cell types. Finally, we summarize the clinical implications of POT1 alterations in the germline and in cancer, and possible approaches for the development of targeted cancer therapies.

The ultrastructural development of the schistosome egg granuloma in mice

Parasitology ◽  
1977 ◽  
Vol 75 (1) ◽  
pp. 119-123 ◽  
Author(s):  
Mary D. Smith
Keyword(s):  
Electron Microscopy ◽  
Schistosoma Mansoni ◽  
Cell Types ◽  
Delayed Hypersensitivity ◽  
Hypersensitivity Response ◽  
Type Reaction ◽  
Early Stages ◽  
Hepatic Granulomas ◽  
Different Cell Types

Hepatic granulomas from mice infected with Schistosoma mansoni for periods of 8 weeks to 1 year were studied by electron microscopy. The different cell types present in the granulomas suggested that whilst a delayed hypersensitivity response predominated during early stages of infection an Arthus-type reaction associated with delayed hypersensi-tivity occurred at later stages of infection.

High-resolution SEM of kidney tissue using cryo-techniques

1990 ◽  
Vol 48 (3) ◽  
pp. 8-9
Author(s):  
P Walther ◽  
P Herter ◽  
J Hentschel ◽  
H Hentschel
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Room Temperature ◽  
Kidney Tissue ◽  
Cell Types ◽  
Electron Gun ◽  
Precise Localization ◽  
Transmission Electron ◽  
Different Cell Types ◽  
Scanning Electron

The kidney is a complex zonated organ with a variety of different cell types. For the study of the functional and morphological features, the precise localization in the zones is relevant, which requires the evaluation of rather large portions of tissue. Transmission electron microscopy of replicas of tissue is limited by difficulties to obtain sufficiently large specimens. In order to overcome this problem cryopreparation methods and high resolution field emission scanning electron microscopy (SEM) were used.1 mm3 cubes of perfusion fixed rabbit kidneys cryoprotected with glycerol were frozen by plunging into liquid propane. For further preparation two different methods were employed.1: Samples were fractured in liquid nitrogen with a scalpel, freeze substituted using methanol with glutaraldehyde and osmiumtetroxid, warmed to room temperature, critical point dried, and coated by electron gun evaporation with 2 nm of platinum at an angle of 45°, and 10 nm of carbon perpendicularly.

Electron-microscopic study of the spindle and chromosome movement in the yeast Saccharomyces cerevisiae

1976 ◽  
Vol 22 (2) ◽  
pp. 219-242 ◽  
Author(s):  
J.B. Peterson ◽  
H. Ris
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Spindle Pole ◽  
Linkage Groups ◽  
Electron Microscopic ◽  
Yeast Saccharomyces Cerevisiae ◽  
Genetic Studies ◽  
Voltage Electron ◽  
High Voltage Electron Microscope ◽  
Spindle Microtubules ◽  
Diploid Cells

Mitosis in yeast Saccharomyces cerevisiae was investigated in thick (0-25-I mum) serial sections with a high voltage electron microscope and in preparations of spheroplasts spread on a water surface. Spindle microtubules originate from a plaque-like structure called the spindle pole bosis the SPB duplicates and a set of long and short microtubules develops on each SPB. The spindle arises as the SPBs separate on the nuclear membrane adense and are not individually visible. Genetic studies, however, have indicated that there are 17 linkage groups. The number of microtubules was determined in diploid and haploid spindles on serial stereo micrographs. In diploid mitosis about 40 microtubules issue from a SPB. Most are non-continuous and often they are visibly associated with a chromatin fibre. The spindle in haploid cells is similar except that the number of microtubules is about half that in diploid cells and the SPB is smaller. The pole-to-pole microtubules vary in number from spindle to spindle, but in each case enough microtubules are present to account for each linkage group being associated with a single non-continuous microtubule. We conclude that mitosis in yeast is comparable in its general aspect to that observed in typical eukaryotes.

scholarly journals Isolation of DNA sequences preferentially expressed during sporulation in Saccharomyces cerevisiae.

1984 ◽  
Vol 4 (1) ◽  
pp. 142-150 ◽  
Author(s):  
A Percival-Smith ◽  
J Segall
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Sequences ◽  
Vegetative Growth ◽  
Cdna Probe ◽  
Cell Types ◽  
Sporulation Medium ◽  
Alpha Cells ◽  
Yeast Saccharomyces Cerevisiae ◽  
Dna Library ◽  
Vector Pbr322

A differential hybridization screen has been used to identify genes cloned from the yeast Saccharomyces cerevisiae that are expressed preferentially during sporulation. Duplicate copies of a partial Sau3A yeast DNA library prepared in the vector pBR322 were hybridized with radioactive cDNA probes representing the mRNA populations of sporulating a alpha cells and asporogenous alpha alpha cells at various times after transfer to sporulation medium. Thirty-eight clones showed an enhanced hybridization signal with the a alpha sporulation probe relative to the alpha alpha control cDNA probe. A comparison of the array of fragments produced by restriction endonuclease digestion of these plasmids suggested that 15 different sequences had been cloned. An RNA blot analysis using these cloned DNAs to probe RNAs purified from aa, a alpha, and alpha alpha cells harvested either during vegetative growth or at 10 h after transfer to sporulation medium indicated that 14 different sporulation-specific genes had been identified. Transcripts complementary to these genes are present only in a alpha cells after transfer to sporulation medium. Three of these clones contain two sporulation-specific genes. Three genes have been identified that are expressed in all cell types during vegetative growth and only in a alpha cells in sporulation medium.

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