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1.
Materials (Basel) ; 16(20)2023 Oct 21.
Article in English | MEDLINE | ID: mdl-37895778

ABSTRACT

Incineration is currently the standard way of disposing of municipal waste. It uses components protected by high-temperature-resistant layers of materials, such as Inconel alloys. Therefore, the objective of the current paper is to study the mechanical properties and structure of a bimetallic Inconel 625-16Mo3 steel tube. The Inconel 625 layer was 3.5 mm thick and was applied to the surface of the tube with a wall thickness of 7 mm via the cold metal transfer method. The bimetallic tube was bent using a supercritical bend (d ≤ 0.7D). This paper is focused on the investigation of the material changes in the Inconel 625 layer areas influenced by the maximum tensile and compressive stresses after the bend. The change in layer thickness after the bend was evaluated and compared to the non-deformed tube. In addition, the local mechanical properties (nanohardness, Young modulus) across the indicated interfacial areas using quasistatic nanoindentation were investigated. Subsequently, a thorough microstructure observation was carried out in areas with maximum tensile and compressive stresses to determine changes in the morphology and size of dendrites related to the effect of tensile or compressive stresses induced by bending. It was found that the grain featured a stretched secondary dendrite axis in the area of tensile stress, but compressive stress imparted a prolongation of the primary dendrite axis.

2.
Front Plant Sci ; 14: 1226498, 2023.
Article in English | MEDLINE | ID: mdl-37636115

ABSTRACT

The plant endomembrane system is an elaborate collection of membrane-bound compartments that perform distinct tasks in plant growth and development, and in responses to abiotic and biotic stresses. Most plant viruses are positive-strand RNA viruses that remodel the host endomembrane system to establish intricate replication compartments. Their fundamental role is to create optimal conditions for viral replication, and to protect replication complexes and the cell-to-cell movement machinery from host defenses. In addition to the intracellular antiviral defense, represented mainly by RNA interference and effector-triggered immunity, recent findings indicate that plant antiviral immunity also includes membrane-localized receptor-like kinases that detect viral molecular patterns and trigger immune responses, which are similar to those observed for bacterial and fungal pathogens. Another recently identified part of plant antiviral defenses is executed by selective autophagy that mediates a specific degradation of viral proteins, resulting in an infection arrest. In a perpetual tug-of-war, certain host autophagy components may be exploited by viral proteins to support or protect an effective viral replication. In this review, we present recent advances in the understanding of the molecular interplay between viral components and plant endomembrane-associated pathways.

3.
Sci Rep ; 13(1): 9233, 2023 Jun 07.
Article in English | MEDLINE | ID: mdl-37286666

ABSTRACT

Drilling into the solid material is one of the basic technological operations, which creates a cylindrical hole in an appropriate time with required quality. Drilling operation demands a favourable removal of chips from the cutting area because a creation of an undesirable shape of chips can impart a lower quality of the drilled hole corresponding with the generation of excess heat due to the intense contact of the chip with drill. The solution for a proper machining is a suitable modification of the drill geometry i.e., point and clearance angles as presented in current study. The tested drills are made of M35 high-speed steel characterized by a very thin core at the point of the drill. An interesting feature of the drills is the use of cutting speed higher than 30 m min-1, with the feed of 0.2 mm per revolution. The surface roughness (Ra and Rz lower than 1 µm and 6 µm respectively), cylindricity (0.045 mm), roundness (0.025 mm), perpendicularity of the hole axis (0.025 mm), diameters and position of the individual holes were achieved for a drill with point angle 138.32°and clearance angle 6.92 respectively. The increase of the drill point angle by 6° resulted in the decrease in the feed force of more than 150 N. In addition, an increase of the clearance angle by 1° resulted with a decrease in the feed force of 70 N. The results of the experiment showed that with the correct geometry of the tool the effective machining without using internal cooling can be realised.

4.
Materials (Basel) ; 16(8)2023 Apr 21.
Article in English | MEDLINE | ID: mdl-37110105

ABSTRACT

The Fused Filament Fabrication (FFF) method is an additive technology that is used for the creation of prototypes within Rapid Prototyping (RP) as well as for the creation of final components in piece or small-series production. The possibility of using FFF technology in the creation of final products requires knowledge of the properties of the material and, at the same time, how these properties change due to degradation effects. In this study, the mechanical properties of the selected materials (PLA, PETG, ABS, and ASA) were tested in their non-degenerate state and after exposure of the samples to the selected degradation factors. For the analysis, which was carried out by the tensile test and the Shore D hardness test, samples of normalized shape were prepared. The effects of UV radiation, high temperature environments, high humidity environments, temperature cycles, and exposure to weather conditions were monitored. The parameters obtained from the tests (tensile strength and Shore D hardness) were statistically evaluated, and the influence of degradation factors on the properties of individual materials was assessed. The results showed that even between individual manufacturers of the same filament there are differences, both in the mechanical properties and in the behavior of the material after exposure to degradation effects.

5.
Eur J Cell Biol ; 102(2): 151309, 2023 Jun.
Article in English | MEDLINE | ID: mdl-36933283

ABSTRACT

The study of endomembrane trafficking is crucial for understanding how cells and whole organisms function. Moreover, there is a special interest in investigating endomembrane trafficking in plants, given its role in transport and accumulation of seed storage proteins and in secretion of cell wall material, arguably the two most essential commodities obtained from crops. The mechanisms of anterograde transport in the biosynthetic and endocytic pathways of plants have been thoroughly discussed in recent reviews, but, comparatively, retrograde trafficking pathways have received less attention. Retrograde trafficking is essential to recover membranes, retrieve proteins that have escaped from their intended localization, maintain homeostasis in maturing compartments, and recycle trafficking machinery for its reuse in anterograde transport reactions. Here, we review the current understanding on retrograde trafficking pathways in the endomembrane system of plants, discussing their integration with anterograde transport routes, describing conserved and plant-specific retrieval mechanisms at play, highlighting contentious issues and identifying open questions for future research.


Subject(s)
Plants , Biological Transport , Plants/metabolism , Protein Transport
6.
Materials (Basel) ; 16(3)2023 Jan 20.
Article in English | MEDLINE | ID: mdl-36769985

ABSTRACT

This paper deals with the study of high-strength M300 maraging steel produced using the selective laser melting method. Heat treatment consists of solution annealing and subsequent aging; the influence of the selected aging temperatures on the final mechanical properties-microhardness and compressive yield strength-and the structure of the maraging steel are described in detail. The microstructure of the samples is examined using optical and electron microscopy. The compressive test results show that the compressive yield strength increased after heat treatment up to a treatment temperature of 480 °C and then gradually decreased. The sample aged at 480 °C also exhibited the highest observed microhardness of 562 HV. The structure of this sample changed from the original melt pools to a relatively fine-grained structure with a high fraction of high-angle grain boundaries (72%).

8.
Polymers (Basel) ; 14(9)2022 Apr 26.
Article in English | MEDLINE | ID: mdl-35566934

ABSTRACT

Within the herein presented research, we studied the applicability of flax fabrics for composite parts in personal watercrafts in order to enhance damping of vibrations from the engine and noise reduction (which is relatively high for contemporary carbon constructions). Since the composite parts are intended to be exposed to humid environments requiring high levels of mechanical properties, a carbon-flax composite was selected. Samples of carbon, fiberglass, flax, and hybrid carbon-flax twill and biax fabrics were subjected to tensile and three-point bending tests. The mechanical properties were also tested after exposure of the samples to a humid environment. Damping was assessed by vibration and noise measurements directly on the complete float for samples as well as real parts. The hybrid carbon-flax material exhibited lower values of tensile strength than the carbon material (760 MPa compared to 463 MPa), but, at the same time, significantly higher than the other tested materials, or flax itself (115 MPa for a twill fabric). A similar trend in the results was observed for the three-point bending tests. Vibration tests and noise measurements showed reductions in vibration amplitude and frequency when using the carbon-flax hybrid material; the frequency response function for the watercraft part assembled from the hybrid material was 50% lower than for that made of carbon. Testing of samples located in a humid environment showed the necessity of surface treatment to prevent moisture absorption (mechanical properties were reduced at minimum by 28%). The tests confirmed that the hybrid material is satisfactory in terms of strength and its contribution to noise and vibration damping.

9.
Proc Natl Acad Sci U S A ; 119(20): e2200492119, 2022 05 17.
Article in English | MEDLINE | ID: mdl-35533279

ABSTRACT

Vacuolar proteins play essential roles in plant physiology and development, but the factors and the machinery regulating their vesicle trafficking through the endomembrane compartments remain largely unknown. We and others have recently identified an evolutionarily conserved plant endosomal sorting complex required for transport (ESCRT)-associated protein apoptosis-linked gene-2 interacting protein X (ALIX), which plays canonical functions in the biogenesis of the multivesicular body/prevacuolar compartment (MVB/PVC) and in the sorting of ubiquitinated membrane proteins. In this study, we elucidate the roles and underlying mechanism of ALIX in regulating vacuolar transport of soluble proteins, beyond its conventional ESCRT function in eukaryotic cells. We show that ALIX colocalizes and physically interacts with the retromer core subunits Vps26 and Vps29 in planta. Moreover, double-mutant analysis reveals the genetic interaction of ALIX with Vps26 and Vps29 for regulating trafficking of soluble vacuolar proteins. Interestingly, depletion of ALIX perturbs membrane recruitment of Vps26 and Vps29 and alters the endosomal localization of vacuolar sorting receptors (VSRs). Taken together, ALIX functions as a unique retromer core subcomplex regulator by orchestrating receptor-mediated vacuolar sorting of soluble proteins.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Carrier Proteins/metabolism , Endosomal Sorting Complexes Required for Transport/metabolism , Endosomes/metabolism , Plants/metabolism , Protein Transport/physiology , Vacuoles/metabolism
10.
Sci Rep ; 12(1): 8645, 2022 05 23.
Article in English | MEDLINE | ID: mdl-35606512

ABSTRACT

Historical field systems are an essential part of the traditional cultural landscape of societies with primarily agricultural subsistence. They embody many functions and values, as they affect the productional, ecological and hydrological functioning of the landscape, its cultural values, the way people perceive the landscape, and their impact on present-day farming. As an aspect of the historical landscape, field systems are a topic investigated in landscape archaeology, environmental studies, historical geography, landscape ecology, and related disciplines. Historical field systems can form many complex spatial structures, shapes and patterns. This paper focuses on identifying environmental and historical/cultural driving forces during the formation and the historical development of various field pattern types. We worked with 523 settlements established in the medieval to the early modern period (approx. 900-1600 AD) in the present-day Czech Republic. We have determined the proportions of different field pattern types in the examined cadastres and have statistically compared them with a variety of environmental and geographical predictors. Our results indicate a strong influence of environmental predictors (terrain undulation, cadastre size), the impact of specific historical events and associated social changes (e.g. land confiscations by the state in the seventeenth century), and a significant relationship between field pattern types and settlement layout types. Furthermore, we have observed the different adaptations of field pattern types to similar environmental conditions, as well as the impact of social and political factors on the processes of landscape formation. Our paper provides the first detailed analysis of the geographical distribution of traditional field systems on the scale of an entire modern state, and emphasizes the importance of transdisciplinary research on cultural landscapes.


Subject(s)
Agriculture , Archaeology , Conservation of Natural Resources , Czech Republic , Ecosystem , Farms , Geography , Humans
11.
Proc Natl Acad Sci U S A ; 117(18): 9884-9895, 2020 05 05.
Article in English | MEDLINE | ID: mdl-32321832

ABSTRACT

The factors and mechanisms involved in vacuolar transport in plants, and in particular those directing vesicles to their target endomembrane compartment, remain largely unknown. To identify components of the vacuolar trafficking machinery, we searched for Arabidopsis modified transport to the vacuole (mtv) mutants that abnormally secrete the synthetic vacuolar cargo VAC2. We report here on the identification of 17 mtv mutations, corresponding to mutant alleles of MTV2/VSR4, MTV3/PTEN2A MTV7/EREL1, MTV8/ARFC1, MTV9/PUF2, MTV10/VPS3, MTV11/VPS15, MTV12/GRV2, MTV14/GFS10, MTV15/BET11, MTV16/VPS51, MTV17/VPS54, and MTV18/VSR1 Eight of the MTV proteins localize at the interface between the trans-Golgi network (TGN) and the multivesicular bodies (MVBs), supporting that the trafficking step between these compartments is essential for segregating vacuolar proteins from those destined for secretion. Importantly, the GARP tethering complex subunits MTV16/VPS51 and MTV17/VPS54 were found at endoplasmic reticulum (ER)- and microtubule-associated compartments (EMACs). Moreover, MTV16/VPS51 interacts with the motor domain of kinesins, suggesting that, in addition to tethering vesicles, the GARP complex may regulate the motors that transport them. Our findings unveil a previously uncharacterized compartment of the plant vacuolar trafficking pathway and support a role for microtubules and kinesins in GARP-dependent transport of soluble vacuolar cargo in plants.


Subject(s)
Arabidopsis Proteins/metabolism , Protein Transport/genetics , Vacuoles/metabolism , Vesicular Transport Proteins/genetics , Alleles , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Cytoplasmic Vesicles/genetics , Cytoplasmic Vesicles/metabolism , Endoplasmic Reticulum/genetics , Endoplasmic Reticulum/metabolism , Golgi Apparatus/genetics , Golgi Apparatus/metabolism , Kinesins/genetics , Kinesins/metabolism , Microtubules/genetics , Microtubules/metabolism , Multivesicular Bodies/genetics , Multivesicular Bodies/metabolism , Mutation , Vacuoles/genetics , Vesicular Transport Proteins/metabolism
12.
Plant J ; 97(5): 805-824, 2019 03.
Article in English | MEDLINE | ID: mdl-30748050

ABSTRACT

The phytohormone cytokinin has been shown to affect many aspects of plant development ranging from the regulation of the shoot apical meristem to leaf senescence. However, some studies have reported contradictory effects of cytokinin on leaf physiology. Therefore cytokinin treatments cause both chlorosis and increased greening and both lead to decrease or increase in cell size. To elucidate this multifaceted role of cytokinin in leaf development, we have employed a system of temporal controls over the cytokinin pool and investigated the consequences of modulated cytokinin levels in the third leaf of Arabidopsis. We show that, at the cell proliferation phase, cytokinin is needed to maintain cell proliferation by blocking the transition to cell expansion and the onset of photosynthesis. Transcriptome profiling revealed regulation by cytokinin of a gene suite previously shown to affect cell proliferation and expansion and thereby a molecular mechanism by which cytokinin modulates a molecular network underlying the cellular responses. During the cell expansion phase, cytokinin stimulates cell expansion and differentiation. Consequently, a cytokinin excess at the cell expansion phase results in an increased leaf and rosette size fueled by higher cell expansion rate, yielding higher shoot biomass. Proteome profiling revealed the stimulation of primary metabolism by cytokinin, in line with an increased sugar content that is expected to increase turgor pressure, representing the driving force of cell expansion. Therefore, the developmental timing of cytokinin content fluctuations, together with a tight control of primary metabolism, is a key factor mediating transitions from cell proliferation to cell expansion in leaves.


Subject(s)
Arabidopsis/physiology , Cytokinins/metabolism , Plant Growth Regulators/metabolism , Proteome , Signal Transduction , Transcriptome , Arabidopsis/genetics , Arabidopsis/growth & development , Cell Enlargement , Cell Proliferation , Gene Ontology , Plant Leaves/genetics , Plant Leaves/growth & development , Plant Leaves/physiology
13.
Proc Biol Sci ; 285(1871)2018 01 31.
Article in English | MEDLINE | ID: mdl-29386368

ABSTRACT

Human populations tend to grow steadily, because of the ability of people to make innovations, and thus overcome and extend the limits imposed by natural resources. It is therefore questionable whether traditional concepts of population ecology, including environmental carrying capacity, can be applied to human societies. The existence of carrying capacity cannot be simply inferred from population time-series, but it can be indicated by the tendency of populations to return to a previous state after a disturbance. So far only indirect evidence at a coarse-grained scale has indicated the historical existence of human carrying capacity. We analysed unique historical population data on 88 settlements before and after the Thirty Years War (1618-1648), one the longest and most destructive conflicts in European history, which reduced the population of Central Europe by 30-50%. The recovery rate of individual settlements after the war was positively correlated with the extent of the disturbance, so that the population size of the settlements after a period of regeneration was similar to the pre-war situation, indicating an equilibrium population size (i.e. carrying capacity). The carrying capacity of individual settlements was positively determined mostly by the fertility of the soil and the area of the cadastre, and negatively by the number of other settlements in the surroundings. Pre-industrial human population sizes were thus probably controlled by negative density dependence mediated by soil fertility, which could not increase due to limited agricultural technologies.


Subject(s)
Conservation of Natural Resources , Population Dynamics , Rural Population/statistics & numerical data , Agriculture/instrumentation , Czech Republic , History, 17th Century , History, 18th Century , Humans , Technology
14.
Methods Mol Biol ; 1511: 113-118, 2017.
Article in English | MEDLINE | ID: mdl-27730606

ABSTRACT

Isolation of various subcellular compartments followed by a high-coverage proteomic analysis provides an unparalleled foundation for the functional analyses of proteins. Analyses of tonoplast preparations free of major contaminants provide insights into vesicular fusion machinery, solute transport, and the vacuole association with the cytoskeleton, whereas analyses of the vacuolar lumen have yielded numerous soluble glycosidases, proteases, and proteins involved in stress responses. In addition, vacuolar lumen preparations have also allowed characterization of a luminal solute content in response to various abiotic stresses. Here, I revisit and update one of the most successful methodologies for vacuole and tonoplast isolation.


Subject(s)
Arabidopsis/chemistry , Cell Fractionation/methods , Intracellular Membranes/chemistry , Plant Leaves/chemistry , Vacuoles/chemistry , Arabidopsis/growth & development , Cell Fractionation/instrumentation , Centrifugation, Density Gradient/instrumentation , Centrifugation, Density Gradient/methods , Culture Media/chemistry , Plant Leaves/growth & development , Povidone/chemistry , Protoplasts/chemistry , Silicon Dioxide/chemistry
15.
Front Plant Sci ; 7: 650, 2016.
Article in English | MEDLINE | ID: mdl-27242851

ABSTRACT

Unfavorable environmental and developmental conditions may cause disturbances in protein folding in the endoplasmic reticulum (ER) that are recognized and counteracted by components of the Unfolded Protein Response (UPR) signaling pathways. The early cellular responses include transcriptional changes to increase the folding and processing capacity of the ER. In this study, we systematically screened a collection of inducible transgenic Arabidopsis plants expressing a library of transcription factors for resistance toward UPR-inducing chemicals. We identified 23 candidate genes that may function as novel regulators of the UPR and of which only three genes (bZIP10, TBF1, and NF-YB3) were previously associated with the UPR. The putative role of identified candidate genes in the UPR signaling is supported by favorable expression patterns in both developmental and stress transcriptional analyses. We demonstrated that WRKY75 is a genuine regulator of the ER-stress cellular responses as its expression was found to be directly responding to ER stress-inducing chemicals. In addition, transgenic Arabidopsis plants expressing WRKY75 showed resistance toward salt stress, connecting abiotic and ER-stress responses.

16.
Zoo Biol ; 35(2): 147-56, 2016.
Article in English | MEDLINE | ID: mdl-26741794

ABSTRACT

Almost nothing is known about the natural vocal behavior of lesser galagos living in zoos. This is perhaps because they are usually kept in nocturnal exhibits separated from the visitors by a transparent and acoustically insulating glass barrier. The aim of the present study was therefore to fill this gap in knowledge of the vocal behavior of lesser galagos from zoos. This knowledge might be beneficial because the vocalizations of these small primates can be used for species determination. We performed a 10-day-long acoustic monitoring of vocal activity in each of seven various groups of Galago senegalensis and G. moholi living at four zoos. We quantitatively evaluated the occurrence of four loud vocalization types present in both species, including the most species-specific advertisement call. We found that qualitative as well as quantitative differences exist in the vocal behavior of the studied groups. We confirmed that the observed vocalization types can be collected from lesser galagos living at zoos, and the success can be increased by selecting larger and more diverse groups. We found two distinct patterns of diel vocal activity in the most vocally active groups. G. senegalensis groups were most vocally active at the beginning and at the end of their activity period, whereas one G. moholi group showed an opposite pattern. The latter is surprising, as it is generally accepted that lesser galagos emit advertisement calls especially at dawn and dusk, i.e., at the beginning and at the end of their diel activity.


Subject(s)
Animals, Zoo/physiology , Galago/physiology , Vocalization, Animal/physiology , Animals
17.
Zoo Biol ; 34(6): 503-12, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26152313

ABSTRACT

The European ground squirrel (Spermophilus citellus) emits alarm calls that warn conspecifics of potential danger. Although it has been observed that inexperienced juveniles of this species emit alarm calls that sound similar to those of adults, studies focusing on juvenile alarm calls are lacking. We analyzed the acoustic structure of alarm calls emitted by six permanently marked European ground squirrels living in a semi-natural enclosure when they were juveniles and after 1 year as adults. We found that the acoustic structure of the juvenile alarm calls was significantly different from those of adults and that the alarm calls underwent nearly the same changes in all studied individuals. All juveniles emitted alarm calls consisting of one element with almost constant frequency, but their alarm calls included a second frequency-modulated element after their first hibernation as adults. Our data show that the duration of the first element is significantly shorter in adults than in juveniles. Additionally, the frequency of the first element is significantly higher in adults than in juveniles. Similar to previous findings in other Palearctic ground squirrel species, our data are inconsistent with the assumption that juvenile mammals emit vocalizations with higher fundamental frequencies than adults. However, our results do not support the previously suggested hypothesis that juvenile ground squirrels conceal information regarding their age in their alarm calls because we found significant differences in alarm calls of juveniles and adults.


Subject(s)
Animals, Zoo/physiology , Sciuridae/physiology , Vocalization, Animal/physiology , Acoustics , Age Factors , Animals , Hibernation , Housing, Animal/standards
18.
Biol Res ; 48: 39, 2015 Jul 25.
Article in English | MEDLINE | ID: mdl-26209329

ABSTRACT

BACKGROUND: A highly regulated trafficking of cargo vesicles in eukaryotes performs protein delivery to a variety of cellular compartments of endomembrane system. The two main routes, the secretory and the endocytic pathways have pivotal functions in uni- and multi-cellular organisms. Protein delivery and targeting includes cargo recognition, vesicle formation and fusion. Developing new tools to modulate protein trafficking allows better understanding the endomembrane system mechanisms and their regulation. The compound Sortin2 has been described as a protein trafficking modulator affecting targeting of the vacuolar protein carboxypeptidase Y (CPY), triggering its secretion in Saccharomyces cerevisiae. RESULTS: A reverse chemical-genetics approach was used to identify key proteins for Sortin2 bioactivity. A genome-wide Sortin2 resistance screen revealed six yeast deletion mutants that do not secrete CPY when grown at Sortin2 condition where the parental strain does: met18, sla1, clc1, dfg10, dpl1 and yjl175w. Integrating mutant phenotype and gene ontology annotation of the corresponding genes and their interactome pointed towards a high representation of genes involved in the endocytic process. In wild type yeast endocytosis towards the vacuole was faster in presence of Sortin2, which further validates the data of the genome-wide screen. This effect of Sortin2 depends on structural features of the molecule, suggesting compound specificity. Sortin2 did not affect endocytic trafficking in Sortin2-resistant mutants, strongly suggesting that the Sortin2 effects on the secretory and endocytic pathways are linked. CONCLUSIONS: Overall, the results reveal that Sortin2 enhances the endocytic transport pathway in Saccharomyces cerevisiae. This cellular effect is most likely at the level where secretory and endocytic pathways are merged. Them Sortin2 specificity over the endomembrane system places it as a powerful biological modulator for cell biology.


Subject(s)
Alkanesulfonates/pharmacology , Endocytosis/physiology , Plant Proteins/physiology , Protein Transport , Rhodanine/analogs & derivatives , Saccharomyces cerevisiae/metabolism , Vacuoles/metabolism , Biological Transport , Phenotype , Protein Transport/genetics , Rhodanine/pharmacology , Secretory Pathway , Vacuoles/physiology
19.
Biol. Res ; 48: 1-11, 2015. ilus, graf, tab
Article in English | LILACS | ID: biblio-950803

ABSTRACT

BACKGROUND: A highly regulated trafficking of cargo vesicles in eukaryotes performs protein delivery to a variety of cellular compartments of endomembrane system. The two main routes, the secretory and the endocytic pathways have pivotal functions in uni- and multi-cellular organisms. Protein delivery and targeting includes cargo recognition, vesicle formation and fusion. Developing new tools to modulate protein trafficking allows better understanding the endomembrane system mechanisms and their regulation. The compound Sortin2 has been described as a protein trafficking modulator affecting targeting of the vacuolar protein carboxypeptidase Y (CPY), triggering its secretion in Saccharomyces cerevisiae. RESULTS: A reverse chemical-genetics approach was used to identify key proteins for Sortin2 bioactivity. A genome-wide Sortin2 resistance screen revealed six yeast deletion mutants that do not secrete CPY when grown at Sortin2 condition where the parental strain does: met18, sla1, clc1, dfg10, dpl1 and yjl175w. Integrating mutant phenotype and gene ontology annotation of the corresponding genes and their interactome pointed towards a high representation of genes involved in the endocytic process. In wild type yeast endocytosis towards the vacuole was faster in presence of Sortin2, which further validates the data of the genome-wide screen. This effect of Sortin2 depends on structural features of the molecule, suggesting compound specificity. Sortin2 did not affect endocytic trafficking in Sortin2-resistant mutants, strongly suggesting that the Sortin2 effects on the secretory and endocytic pathways are linked. CONCLUSIONS: Overall, the results reveal that Sortin2 enhances the endocytic transport pathway in Saccharomyces cerevisiae. This cellular effect is most likely at the level where secretory and endocytic pathways are merged. Them Sortin2 specificity over the endomembrane system places it as a powerful biological modulator for cell biology.


Subject(s)
Plant Proteins/physiology , Rhodanine/analogs & derivatives , Saccharomyces cerevisiae/metabolism , Vacuoles/metabolism , Alkanesulfonates/pharmacology , Protein Transport/genetics , Endocytosis/physiology , Phenotype , Rhodanine/pharmacology , Vacuoles/physiology , Biological Transport , Secretory Pathway
20.
PLoS One ; 9(11): e111571, 2014.
Article in English | MEDLINE | ID: mdl-25390304

ABSTRACT

Shrews have rich vocal repertoires that include vocalizations within the human audible frequency range and ultrasonic vocalizations. Here, we recorded and analyzed in detail the acoustic structure of a vocalization with unclear functional significance that was spontaneously produced by 15 adult, captive Asian house shrews (Suncus murinus) while they were lying motionless and resting in their nests. This vocalization was usually emitted repeatedly in a long series with regular intervals. It showed some structural variability; however, the shrews most frequently emitted a tonal, low-frequency vocalization with minimal frequency modulation and a low, non-vocal click that was clearly noticeable at its beginning. There was no effect of sex, but the acoustic structure of the analyzed vocalizations differed significantly between individual shrews. The encoded individuality was low, but it cannot be excluded that this individuality would allow discrimination of family members, i.e., a male and female with their young, collectively resting in a common nest. The question remains whether the Asian house shrews indeed perceive the presence of their mates, parents or young resting in a common nest via the resting-associated vocalization and whether they use it to discriminate among their family members. Additional studies are needed to explain the possible functional significance of resting-associated vocalizations emitted by captive Asian house shrews. Our study highlights that the acoustic communication of shrews is a relatively understudied topic, particularly considering that they are highly vocal mammals.


Subject(s)
Acoustics , Shrews/physiology , Vocalization, Animal , Animals , Body Weight , Female , Male , Rest , Social Behavior
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