scholarly journals Generation of an Isogenic Collection of Yeast Actin Mutants and Identification of Three Interrelated Phenotypes

Genetics ◽  
2001 ◽  
Vol 157 (2) ◽  
pp. 533-543
Author(s):  
Johanna L Whitacre ◽  
Dana A Davis ◽  
Kurt A Toenjes ◽  
Sharon M Brower ◽  
Alison E M Adams
Keyword(s):  
Crystal Structure ◽  
Growth Rates ◽  
Small Region ◽  
Wild Type ◽  
Large Collection ◽  
Growth Defects ◽  
Cytoskeletal Function ◽  
Highly Correlated ◽  
The Relationship ◽  
Mutant Cells

Abstract A large collection of yeast actin mutations has been previously isolated and used in numerous studies of actin cytoskeletal function. However, the various mutations have been in congenic, rather than isogenic, backgrounds, making it difficult to compare the subtle phenotypes that are characteristic of these mutants. We have therefore placed 27 mutations in an isogenic background. We used a subset of these mutants to compare the degree to which different actin alleles are defective in sporulation, endocytosis, and growth on NaCl-containing media. We found that the three phenotypes are highly correlated. The correlations are specific and not merely a reflection of general growth defects, because the phenotypes are not correlated with growth rates under normal conditions. Significantly, those actin mutants exhibiting the most severe phenotypes in all three processes have altered residues that cluster to a small region of the actin crystal structure previously defined as the fimbrin (Sac6p)-binding site. We examined the relationship between endocytosis and growth on salt and found that shifting wild-type or actin mutant cells to high salt reduces the rate of α-factor internalization. These results suggest that actin mutants may be unable to grow on salt because of additive endocytic defects (due to mutation and salt).

Plastid Signals Confer Arabidopsis Tolerance to Water Stress

2011 ◽  
Vol 66 (1-2) ◽  
pp. 47-54 ◽  
Author(s):  
Jian Cheng ◽  
Chun-Xia He ◽  
Zhong-Wei Zhang ◽  
Fei Xu ◽  
Da-Wei Zhang ◽  
...  
Keyword(s):  
Water Stress ◽  
Photosynthetic Apparatus ◽  
Nuclear Gene ◽  
Stress Adaptation ◽  
Wild Type ◽  
Oxidative Damages ◽  
Nuclear Gene Expression ◽  
Retrograde Signalling ◽  
The Relationship ◽  
Mutant Cells

Plastid-to-nucleus retrograde signalling coordinates nuclear gene expression with chloroplast function and is essential for the photoautotrophic life-style of plants. The relationship between plastid signalling and water stress response was investigated with genome uncoupled (gun) mutants, gun1, gun3, and gun5, and an abscisic acid (ABA)-responsible transcription factor mutant, abi4. The results showed that gun1, gun3, gun5, and abi4 mutants suffered from more oxidative damages than the wild-type plants under the water stress and the water stress + herbicide (norflurazon, NF) co-treatment. Superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) activities could not be prompted in the plastidsignalling defective mutants under the stress conditions. At the same time, Lhcb expression was not repressed in the plastid-signalling defective mutants by the NF treatment or water stress. Therefore, the photosynthetic apparatus in the mutant cells could not be closed during the stresses and the excessive light caused more photodamages on the mutant leaves. The roles of GUN1, GUN3, GUN5 and ABI4 proteins in environmental stress adaptation have been discussed.

scholarly journals Glc7/Protein Phosphatase 1 Regulatory Subunits Can Oppose the Ipl1/Aurora Protein Kinase by Redistributing Glc7

2006 ◽  
Vol 26 (7) ◽  
pp. 2648-2660 ◽  
Author(s):  
Benjamin A. Pinsky ◽  
Chitra V. Kotwaliwale ◽  
Sean Y. Tatsutani ◽  
Christopher A. Breed ◽  
Sue Biggins
Keyword(s):  
Protein Kinase ◽  
Chromosome Segregation ◽  
Protein Phosphatase ◽  
Kinase Activity ◽  
Budding Yeast ◽  
Protein Phosphatase 1 ◽  
Wild Type ◽  
Regulatory Subunits ◽  
The Relationship ◽  
Mutant Cells

ABSTRACT Faithful chromosome segregation depends on the opposing activities of the budding yeast Glc7/PP1 protein phosphatase and Ipl1/Aurora protein kinase. We explored the relationship between Glc7 and Ipl1 and found that the phosphorylation of the Ipl1 substrate, Dam1, was altered by decreased Glc7 activity, whereas Ipl1 levels, localization, and kinase activity were not. These data strongly suggest that Glc7 ensures accurate chromosome segregation by dephosphorylating Ipl1 targets rather than regulating the Ipl1 kinase. To identify potential Glc7 and Ipl1 substrates, we isolated ipl1-321 dosage suppressors. Seven genes (SDS22, BUD14, GIP3, GIP4, SOL1, SOL2, and PEX31) encode newly identified ipl1 dosage suppressors, and all 10 suppressors encode proteins that physically interact with Glc7. The overexpression of the Gip3 and Gip4 suppressors altered Glc7 localization, indicating they are previously unidentified Glc7 regulatory subunits. In addition, the overexpression of Gip3 and Gip4 from the galactose promoter restored Dam1 phosphorylation in ipl1-321 mutant cells and caused wild-type cells to arrest in metaphase with unsegregated chromosomes, suggesting that Gip3 and Gip4 overexpression impairs Glc7's mitotic functions. We therefore propose that the overexpression of Glc7 regulatory subunits can titrate Glc7 away from relevant Ipl1 targets and thereby suppress ipl1-321 cells by restoring the balance of phosphatase/kinase activity.

Motility and substratum adhesion of Dictyostelium wild-type and cytoskeletal mutant cells: a study by RICM/bright-field double-view image analysis

1995 ◽  
Vol 108 (4) ◽  
pp. 1519-1530 ◽  
Author(s):  
I. Weber ◽  
E. Wallraff ◽  
R. Albrecht ◽  
G. Gerisch
Keyword(s):  
Image Analysis ◽  
Cell Body ◽  
Cytoskeletal Proteins ◽  
Chemotactic Response ◽  
Wild Type ◽  
Bright Field ◽  
Competent Cells ◽  
The Relationship ◽  
Mutant Cells ◽  
Front Region

To investigate the dynamics of cell-substratum adhesion during locomotion, a double-view optical technique and computer-assisted image analysis has been developed which combines reflection interference contrast microscopy (RICM) with bright-field imaging. The simultaneous recording of cell-substratum contact and cell body contour has been applied to aggregation-competent cells of Dictyostelium discoideum. These cells are distinguished from cells at earlier stages of development by small areas of contact to a substratum. Three questions have been addressed in analysing the locomotion of aggregation-competent cells. (1) What is the relationship between changes in the shape of cells and their contact to a substratum during a chemotactic response? (2) What is the relationship between protrusion and retraction of the cell body, and between local attachment and detachment? (3) Are there differences between wild-type and mutant cells that lack certain cytoskeletal proteins? During a chemotactic response the front region of the amoeba can bend towards the gradient of attractant without being supported by its contact with a surface, which excludes the necessity for gradients of adhesion for the response. The finding that in locomoting cells protrusion of the leading edge often precedes retraction establishes a pioneer role for the front region. The finding that gain of contact area precedes loss provides evidence for the coordination of interactions between the cell surface and a substratum. For comparison with wild-type, aggregation-competent triple mutant cells have been used that lack two F-actin crosslinking proteins, alpha-actinin and 120 kDa gelation factor, and an actin filament fragmenting protein, severin. Disturbances in the spatial and temporal control of cytoskeletal activities have been unravelled in the mutant by RICM and quantified by cross-correlation analysis of attachment and detachment vectors. In order to detect these disturbances, it was essential to analyse cell locomotion on the weakly adhesive surface of freshly cleaved mica.

Spatial Variation in Food-Limited Growth of Juvenile Greenback Flounder, Rhombosolea tapirina: Evidence from Otolith Daily Increments and Otolith Scaling

1993 ◽  
Vol 50 (12) ◽  
pp. 2558-2567 ◽  
Author(s):  
Gregory P. Jenkins ◽  
Megan Shaw ◽  
Bryce D. Stewart
Keyword(s):  
Growth Rate ◽  
Growth Rates ◽  
Somatic Growth ◽  
Feeding Rates ◽  
Otolith Growth ◽  
Somatic Growth Rate ◽  
Daily Increment ◽  
Daily Increments ◽  
Highly Correlated ◽  
The Relationship

Growth rates of juvenile flounder, Rhombosolea tapirina, determined from daily increment number, and the relationship between otolith and fish sizes (otolith scaling), were compared between two adjacent areas. Swan Bay, Victoria, a sheltered bay with a well-developed seagrass-detrital system, supports higher populations of prey and feeding rates of juvenile flounder than Port Phillip Bay, an area more exposed to waves and tidal currents. Temperature was significantly higher in Swan Bay (though generally less than 1 °C). Growth rates determined from daily increment number were similar within bays, but significantly different between bays. The pooled growth rate for Swan Bay (0.29 mm∙d−1) was significantly higher than for Port Phillip Bay (0.17 mm∙d−1). The same pattern was found for otolith scaling. Most of the variation in growth rates between the two bays was apparently related to food supply. A laboratory experiment indicated that otolith growth rate had a minimum level which was independent of somatic growth rate, and an additional component which was highly correlated with somatic growth rate. This resulted in an exponential decrease in otolith growth per unit somatic growth with increasing somatic growth rate such that variation in otolith scaling would be greatest at low growth rates.

scholarly journals Asymmetric properties of the Chlamydomonas reinhardtii cytoskeleton direct rhodopsin photoreceptor localization

2011 ◽  
Vol 193 (4) ◽  
pp. 741-753 ◽  
Author(s):  
Telsa M. Mittelmeier ◽  
Joseph S. Boyd ◽  
Mary Rose Lamb ◽  
Carol L. Dieckmann
Keyword(s):  
Chlamydomonas Reinhardtii ◽  
Green Alga ◽  
Basal Body ◽  
Cytoskeletal Protein ◽  
Wild Type ◽  
Unicellular Green Alga ◽  
Protein Mutations ◽  
The Relationship ◽  
Mutant Cells ◽  
In Cells

The eyespot of the unicellular green alga Chlamydomonas reinhardtii is a photoreceptive organelle required for phototaxis. Relative to the anterior flagella, the eyespot is asymmetrically positioned adjacent to the daughter four-membered rootlet (D4), a unique bundle of acetylated microtubules extending from the daughter basal body toward the posterior of the cell. Here, we detail the relationship between the rhodopsin eyespot photoreceptor Channelrhodopsin 1 (ChR1) and acetylated microtubules. In wild-type cells, ChR1 was observed in an equatorial patch adjacent to D4 near the end of the acetylated microtubules and along the D4 rootlet. In cells with cytoskeletal protein mutations, supernumerary ChR1 patches remained adjacent to acetylated microtubules. In mlt1 (multieyed) mutant cells, supernumerary photoreceptor patches were not restricted to the D4 rootlet, and more anterior eyespots correlated with shorter acetylated microtubule rootlets. The data suggest a model in which photoreceptor localization is dependent on microtubule-based trafficking selective for the D4 rootlet, which is perturbed in mlt1 mutant cells.

Ultrastructure and Morphology of the Neurospora Crassa Cell Wall Mutants, Slime And Slime X, Using Tem and Sem

1976 ◽  
Vol 34 ◽  
pp. 54-55
Author(s):  
Karen S. Howard ◽  
H. D. Braymer ◽  
M. D. Socolofsky ◽  
S. A. Milligan
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Wild Type ◽  
Morphological Comparison ◽  
Small Areas ◽  
Solid Media ◽  
Thin Sectioning ◽  
X Cells ◽  
Mutant Cells ◽  
Isolated Cell

The recently isolated cell wall mutant slime X of Neurospora crassa was prepared for ultrastructural and morphological comparison with the cell wall mutant slime. The purpose of this article is to discuss the methods of preparation for TEM and SEM observations, as well as to make a preliminary comparison of the two mutants.TEM: Cells of the slime mutant were prepared for thin sectioning by the method of Bigger, et al. Slime X cells were prepared in the same manner with the following two exceptions: the cells were embedded in 3% agar prior to fixation and the buffered solutions contained 5% sucrose throughout the procedure.SEM: Two methods were used to prepare mutant and wild type Neurospora for the SEM. First, single colonies of mutant cells and small areas of wild type hyphae were cut from solid media and fixed with OSO4 vapors similar to the procedure used by Harris, et al. with one alteration. The cell-containing agar blocks were dehydrated by immersion in 2,2-dimethoxypropane (DMP).

Do Extra-Platelet Sources Contribute to the Plasma Level of Thrombospondin?

1988 ◽  
Vol 59 (02) ◽  
pp. 273-276 ◽  
Author(s):  
J Dawes ◽  
D A Pratt ◽  
M S Dewar ◽  
F E Preston
Keyword(s):  
Platelet Count ◽  
Background Concentration ◽  
Serum Concentrations ◽  
Bone Marrow Depression ◽  
Serial Sampling ◽  
Control Serum ◽  
Platelet Release ◽  
Highly Correlated ◽  
The Relationship

SummaryThrombospondin, a trimeric glycoprotein contained in the platelet α-granules, has been proposed as a marker of in vivo platelet activation. However, it is also synthesised by a range of other cells. The extraplatelet contribution to plasma levels of thrombospondin was therefore estimated by investigating the relationship between plasma thrombospondin levels and platelet count in samples from profoundly thrombocytopenic patients with marrow hypoplasia, using the platelet-specific α-granule protein β-thromboglobulin as control. Serum concentrations of both proteins were highly correlated with platelet count, but while plasma β-thromboglobulin levels and platelet count also correlated, there was no relationship between the number of platelets and thrombospondin concentrations in plasma. Serial sampling of patients recovering from bone marrow depression indicated that the plasma thrombospondin contributed by platelets is superimposed on a background concentration of at least 50 ng/ml probably derived from a non-platelet source, and plasma thrombospondin levels do not simply reflect platelet release.

scholarly journals Suitability Analysis of 17 Probiotic Type Strains of Lactic Acid Bacteria as Starter for Kimchi Fermentation

Foods ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1435
Author(s):  
Hee Seo ◽  
Jae-Han Bae ◽  
Gayun Kim ◽  
Seul-Ah Kim ◽  
Byung Hee Ryu ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Growth Rates ◽  
Principal Component ◽  
Fermented Foods ◽  
Suitability Analysis ◽  
Moderate Growth ◽  
Health Promoting ◽  
Highly Correlated ◽  
Type Strains

The use of probiotic starters can improve the sensory and health-promoting properties of fermented foods. This study aimed to evaluate the suitability of probiotic lactic acid bacteria (LAB) as a starter for kimchi fermentation. Seventeen probiotic type strains were tested for their growth rates, volatile aroma compounds, metabolites, and sensory characteristics of kimchi, and their characteristics were compared to those of Leuconostoc (Le.) mesenteroides DRC 1506, a commercial kimchi starter. Among the tested strains, Limosilactobacillus fermentum, Limosilactobacillus reuteri, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, and Ligilactobacillus salivarius exhibited high or moderate growth rates in simulated kimchi juice (SKJ) at 37 °C and 15 °C. When these five strains were inoculated in kimchi and metabolite profiles were analyzed during fermentation using GC/MS and 1H-NMR, data from the principal component analysis (PCA) showed that L. fermentum and L. reuteri were highly correlated with Le. mesenteroides in concentrations of sugar, mannitol, lactate, acetate, and total volatile compounds. Sensory test results also indicated that these three strains showed similar sensory preferences. In conclusion, L. fermentum and L. reuteri can be considered potential candidates as probiotic starters or cocultures to develop health-promoting kimchi products.

scholarly journals EPHA2-dependent outcompetition of KRASG12D mutant cells by wild-type neighbors in the adult pancreas

2021 ◽  
Author(s):  
William Hill ◽  
Andreas Zaragkoulias ◽  
Beatriz Salvador-Barbero ◽  
Geraint J. Parfitt ◽  
Markella Alatsatianos ◽  
...  
Keyword(s):  
Wild Type ◽  
Mutant Cells

scholarly journals Colony spreading of the gliding bacterium Flavobacterium johnsoniae in the absence of the motility adhesin SprB

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Keiko Sato ◽  
Masami Naya ◽  
Yuri Hatano ◽  
Yoshio Kondo ◽  
Mari Sato ◽  
...  
Keyword(s):  
Soft Agar ◽  
Gliding Motility ◽  
Wild Type ◽  
Initial Cell ◽  
Flavobacterium Johnsoniae ◽  
Seeking Behavior ◽  
Colony Spreading ◽  
Gliding Bacterium ◽  
Mutant Cells ◽  
Scanning Electron

AbstractColony spreading of Flavobacterium johnsoniae is shown to include gliding motility using the cell surface adhesin SprB, and is drastically affected by agar and glucose concentrations. Wild-type (WT) and ΔsprB mutant cells formed nonspreading colonies on soft agar, but spreading dendritic colonies on soft agar containing glucose. In the presence of glucose, an initial cell growth-dependent phase was followed by a secondary SprB-independent, gliding motility-dependent phase. The branching pattern of a ΔsprB colony was less complex than the pattern formed by the WT. Mesoscopic and microstructural information was obtained by atmospheric scanning electron microscopy (ASEM) and transmission EM, respectively. In the growth-dependent phase of WT colonies, dendritic tips spread rapidly by the movement of individual cells. In the following SprB-independent phase, leading tips were extended outwards by the movement of dynamic windmill-like rolling centers, and the lipoproteins were expressed more abundantly. Dark spots in WT cells during the growth-dependent spreading phase were not observed in the SprB-independent phase. Various mutations showed that the lipoproteins and the motility machinery were necessary for SprB-independent spreading. Overall, SprB-independent colony spreading is influenced by the lipoproteins, some of which are involved in the gliding machinery, and medium conditions, which together determine the nutrient-seeking behavior.

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