Assessment of pheromone production and response in fission yeast by a halo test of induced sporulation

Egel, Richard; Willer, Martin; Kjærulff, Søren; Davey, John; Nielsen, Olaf

doi:10.1002/yea.320101012

Cited by 100 publications

(83 citation statements)

References 33 publications

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“…For assessing cells during exponential growth, cells were grown in rich medium (YE), Edinburgh minimal medium (EMM), or minimal sporulation medium with nitrogen (MSL + N) supplemented with amino acids as required. For assessing cells during the mating process, liquid or agar minimal sporulation medium without nitrogen (MSL − N) was used (Egel et al 1994;Vjestica et al 2016). All live-cell imaging was performed on MSL − N agarose pads (Vjestica et al 2016) or in microfluidics chambers (described below).…”

Section: Strains Media and Growth Conditionsmentioning

confidence: 99%

Spatial focalization of pheromone/MAPK signaling triggers commitment to cell–cell fusion

2016

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Cell fusion is universal in eukaryotes for fertilization and development, but what signals this process is unknown. Here, we show in Schizosaccharomyces pombe that fusion does not require a dedicated signal but is triggered by spatial focalization of the same pheromone-GPCR (G-protein-coupled receptor)-MAPK signaling cascade that drives earlier mating events. Autocrine cells expressing the receptor for their own pheromone trigger fusion attempts independently of cell-cell contact by concentrating pheromone release at the fusion focus, a dynamic actin aster underlying the secretion of cell wall hydrolases. Pheromone receptor and MAPK cascade are similarly enriched at the fusion focus, concomitant with fusion commitment in wild-type mating pairs. This focalization promotes cell fusion by immobilizing the fusion focus, thus driving local cell wall dissolution. We propose that fusion commitment is imposed by a local increase in MAPK concentration at the fusion focus, driven by a positive feedback between fusion focus formation and focalization of pheromone release and perception.

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Section: Strains Media and Growth Conditionsmentioning

confidence: 99%

Spatial focalization of pheromone/MAPK signaling triggers commitment to cell–cell fusion

2016

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“…Also used in certain experiments was strain 2694 (h¯ leu1¯ ade6-210 ura4-D18 GFP:Swi6 GFP:atb2), which was a gift from Alison Pidoux (Pidoux et al, 2000). Cells were grown on rich YES or minimal selective media plates at either 25°C or 30°C (Egel et al, 1994;Moreno et al, 1991).…”

Section: Yeast Strainsmentioning

confidence: 99%

“…For microscopy, cells were grown on rich YES or MSA minimal media plates (Egel et al, 1994;Moreno et al, 1991), and a small amount was scraped off plates and resuspended in 20 µl dH2O or 5 µM n-propyl gallate to prevent photobleaching. 1-2 µl of this cell suspension was placed onto an agarose pad (1.1% Apex agarose in PM media) and sealed under a glass coverslip with silicone lubricant vacuum grease (Dow-Corning).…”

Section: Microscopy Of Living Cellsmentioning

confidence: 99%

Individual microtubule dynamics contribute to the function of mitotic and cytoplasmic arrays in fission yeast

Sagolla

Uzawa

Cande

2003

Journal of Cell Science

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Schizosaccharomyces pombe is an excellent organism for studying microtubule dynamics owing to the presence of well-defined microtubule arrays that undergo dramatic rearrangements during various stages of the cell cycle. Using sensitive time-lapse video microscopy and kymographic analysis, we have determined the polymerization/depolymerization kinetics of individual microtubules within these arrays throughout the fission yeast cell cycle. Interphase bundles are composed of 4-7 microtubules that act autonomously, demonstrating that individual microtubules are responsible for mediating the functions ascribed to these arrays. The nucleation and growth of cytoplasmic microtubules is inhibited upon cellular transition into mitosis, leading to their gradual disappearance. At the onset of mitosis, microtubules form on the nuclear face of the spindle pole body and exhibit dramatically increased dynamics. The presence of these intra-nuclear astral microtubules (INA) is reminiscent of spindle assembly and the search and chromosome capture mechanism observed in metazoan cells. Consistent with other in vivo studies, we do not observe microtubule flux in the anaphase B spindle. Finally, the depolymerization of individual microtubules alternates between each half-spindle, resulting in spindle collapse during telophase. On the basis of these observations, we conclude that microtubules in these diverse cytoskeletal arrays have autonomous behaviors that are an essential component of any model describing cell-cycle-dependent changes in the behavior and function of microtubule arrays.

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“…Phenotypes were examined after 22 h of fusion protein expression. The strain myo2-gc was created with the use of the method of Bahler et al (1998b) Genetic crosses to create double mutants with mutants in the SIN pathway were carried as described in Egel et al (1994). The strains used in this article are listed in Table 1.…”

Section: Cell Culture and Strainsmentioning

confidence: 99%

Localization of Fission Yeast Type II Myosin, Myo2, to the Cytokinetic Actin Ring Is Regulated by Phosphorylation of a C-Terminal Coiled-Coil Domain and Requires a Functional Septation Initiation Network

Mulvihill

Barretto

Hyams

2001

MBoC

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Myo2 truncations fused to green fluorescent protein (GFP) defined a C-terminal domain essential for the localization of Myo2 to the cytokinetic actin ring (CAR). The localization domain contained two predicted phosphorylation sites. Mutation of serine 1518 to alanine (S1518A) abolished Myo2 localization, whereas Myo2 with a glutamic acid at this position (S1518E) localized to the CAR. GFP-Myo2 formed rings in the septation initiation kinase (SIN) mutant cdc7-24 at 25°C but not at 36°C. GFP-Myo2S1518E rings persisted at 36°C incdc7-24 but not in another SIN kinase mutant,sid2-250. To further examine the relationship between Myo2 and the SIN pathway, the chromosomal copy ofmyo2 + was fused to GFP (strainmyo2-gc). Myo2 ring formation was abolished in the double mutants myo2-gc cdc7.24 and myo2-gc sid2-250 at the restrictive temperature. In contrast, activation of the SIN pathway in the double mutant myo2-gc cdc16-116 resulted in the formation of Myo2 rings which subsequently collapsed at 36°C. We conclude that the SIN pathway that controls septation in fission yeast also regulates Myo2 ring formation and contraction. Cdc7 and Sid2 are involved in ring formation, in the case of Cdc7 by phosphorylation of a single serine residue in the Myo2 tail. Other kinases and/or phosphatases may control ring contraction.

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Assessment of pheromone production and response in fission yeast by a halo test of induced sporulation

Cited by 100 publications

References 33 publications

Spatial focalization of pheromone/MAPK signaling triggers commitment to cell–cell fusion

Spatial focalization of pheromone/MAPK signaling triggers commitment to cell–cell fusion

Individual microtubule dynamics contribute to the function of mitotic and cytoplasmic arrays in fission yeast

Localization of Fission Yeast Type II Myosin, Myo2, to the Cytokinetic Actin Ring Is Regulated by Phosphorylation of a C-Terminal Coiled-Coil Domain and Requires a Functional Septation Initiation Network

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