scholarly journals Myosin-V, Kinesin-1, and Kinesin-3 Cooperate in Hyphal Growth of the FungusUstilago maydis

2005 ◽  
Vol 16 (11) ◽  
pp. 5191-5201 ◽  
Author(s):  
Isabel Schuchardt ◽  
Daniela Aßmann ◽  
Eckhard Thines ◽  
Christian Schuberth ◽  
Gero Steinberg
Keyword(s):  
Fluorescent Protein ◽  
Hyphal Growth ◽  
Myosin V ◽  
Long Distance ◽  
Long Distance Transport ◽  
Conventional Kinesin ◽  
Green Fluorescent ◽  
Mutant Cells ◽  
Distance Transport

Long-distance transport is crucial for polar-growing cells, such as neurons and fungal hyphae. Kinesins and myosins participate in this process, but their functional interplay is poorly understood. Here, we investigate the role of kinesin motors in hyphal growth of the plant pathogen Ustilago maydis. Although the microtubule plus-ends are directed to the hyphal tip, of all 10 kinesins analyzed, only conventional kinesin (Kinesin-1) and Unc104/Kif1A-like kinesin (Kinesin-3) were up-regulated in hyphae and they are essential for extended hyphal growth. Δkin1 and Δkin3 mutant hyphae grew irregular and remained short, but they were still able to grow polarized. No additional phenotype was detected in Δkin1rkin3 double mutants, but polarity was lost in Δmyo5rkin1 and Δmyo5rkin3 mutant cells, suggesting that kinesins and class V myosin cooperate in hyphal growth. Consistent with such a role in secretion, fusion proteins of green fluorescent protein and Kinesin-1, Myosin-V, and Kinesin-3 accumulate in the apex of hyphae, a region where secretory vesicles cluster to form the fungal Spitzenkörper. Quantitative assays revealed a role of Kin3 in secretion of acid phosphatase, whereas Kin1 was not involved. Our data demonstrate that just two kinesins and at least one myosin support hyphal growth.

scholarly journals Temperature-Dependent Wsm1 and Wsm2 Gene-Specific Blockage of Viral Long-Distance Transport Provides Resistance to Wheat streak mosaic virus and Triticum mosaic virus in Wheat

2016 ◽  
Vol 29 (9) ◽  
pp. 724-738 ◽  
Author(s):  
Satyanarayana Tatineni ◽  
Everlyne N. Wosula ◽  
Melissa Bartels ◽  
Gary L. Hein ◽  
Robert A. Graybosch
Keyword(s):  
Mosaic Virus ◽  
Fluorescent Protein ◽  
Wheat Streak Mosaic Virus ◽  
Viral Resistance ◽  
Wheat Cultivars ◽  
Temperature Dependent ◽  
Long Distance ◽  
Long Distance Transport ◽  
Long Distance Movement ◽  
Distance Transport

Wheat streak mosaic virus (WSMV) and Triticum mosaic virus (TriMV) are economically important viral pathogens of wheat. Wheat cvs. Mace, carrying the Wsm1 gene, is resistant to WSMV and TriMV, and Snowmass, with Wsm2, is resistant to WSMV. Viral resistance in both cultivars is temperature sensitive and is effective at 18°C or below but not at higher temperatures. The underlying mechanisms of viral resistance of Wsm1 and Wsm2, nonallelic single dominant genes, are not known. In this study, we found that fluorescent protein–tagged WSMV and TriMV elicited foci that were approximately similar in number and size at 18 and 24°C, on inoculated leaves of resistant and susceptible wheat cultivars. These data suggest that resistant wheat cultivars at 18°C facilitated efficient cell-to-cell movement. Additionally, WSMV and TriMV efficiently replicated in inoculated leaves of resistant wheat cultivars at 18°C but failed to establish systemic infection, suggesting that Wsm1- and Wsm2-mediated resistance debilitated viral long-distance transport. Furthermore, we found that neither virus was able to enter the leaf sheaths of inoculated leaves or crowns of resistant wheat cultivars at 18°C but both were able to do so at 24°C. Thus, wheat cvs. Mace and Snowmass provide resistance at the long-distance movement stage by specifically blocking virus entry into the vasculature. Taken together, these data suggest that both Wsm1 and Wsm2 genes similarly confer virus resistance by temperature-dependent impairment of viral long-distance movement.

scholarly journals Cse1p Is Involved in Export of Yeast Importin α from the Nucleus

1998 ◽  
Vol 18 (11) ◽  
pp. 6805-6815 ◽  
Author(s):  
Jens Solsbacher ◽  
Patrick Maurer ◽  
F. Ralf Bischoff ◽  
Gabriel Schlenstedt
Keyword(s):  
Nuclear Export ◽  
Fluorescent Protein ◽  
Nuclear Pore ◽  
Wild Type ◽  
Importin Α ◽  
Localization Signal ◽  
Green Fluorescent ◽  
Mutant Cells

ABSTRACT Proteins bearing a nuclear localization signal (NLS) are targeted to the nucleus by the heterodimeric transporter importin. Importin α binds to the NLS and to importin β, which carries it through the nuclear pore complex (NPC). Importin disassembles in the nucleus, evidently by binding of RanGTP to importin β. The importin subunits are exported separately. We investigated the role of Cse1p, theSaccharomyces cerevisiae homologue of human CAS, in nuclear export of Srp1p (yeast importin α). Cse1p is located predominantly in the nucleus but also is present in the cytoplasm and at the NPC. We analyzed the in vivo localization of the importin subunits fused to the green fluorescent protein in wild-type and cse1-1 mutant cells. Srp1p but not importin β accumulated in nuclei ofcse1-1 mutants, which are defective in NLS import but not defective in NLS-independent import pathways. Purified Cse1p binds with high affinity to Srp1p only in the presence of RanGTP. The complex is dissociated by the cytoplasmic RanGTP-binding protein Yrb1p. Combined with the in vivo results, this suggests that a complex containing Srp1p, Cse1p, and RanGTP is exported from the nucleus and is subsequently disassembled in the cytoplasm by Yrb1p. The formation of the trimeric Srp1p-Cse1p-RanGTP complex is inhibited by NLS peptides, indicating that only NLS-free Srp1p will be exported to the cytoplasm.

The Role of Amino Acids in the Long-Distance Transport of La and Y in the Xylem Sap of Tomato

2008 ◽  
Vol 129 (1-3) ◽  
pp. 239-250 ◽  
Author(s):  
Jun-liang Wu ◽  
Zheng-gui Wei ◽  
Hai-yan Zhao ◽  
Hui-xin Li ◽  
Feng Hu
Keyword(s):  
Amino Acids ◽  
Xylem Sap ◽  
Long Distance ◽  
Long Distance Transport ◽  
Distance Transport

scholarly journals Enigmatic clusters of sandstone boulders on plateaus of the Stołowe Mountains (Sudetes, south-west Poland) – their geoheritage and geotouristic value

Geologos ◽  
2021 ◽  
Vol 27 (3) ◽  
pp. 141-155
Author(s):  
Piotr Migoń ◽  
Krzysztof Parzóch
Keyword(s):  
Late Cretaceous ◽  
Sedimentary Rocks ◽  
Long Distance ◽  
Long Distance Transport ◽  
South West ◽  
History Of ◽  
Clastic Sedimentary ◽  
Distance Transport ◽  
Plateau Level

Abstract Among sites of geomorphological interest in the tableland of the Stołowe Mountains, consisting of clastic sedimentary rocks of Late Cretaceous age, are enigmatic occurrences and clusters of sandstone boulders within plateau levels that are underlain by mudstones and marls. These boulders are allochthonous, having been derived from the quartz sandstone beds that support the upper plateau level and stratigraphically are in excess of 50 m above the altitudinal position of the boulders. Topographic conditions preclude long-distance transport from the escarpment slopes; boulders are hypothesised to be the last remnants of completely degraded outliers (mesas) of the upper plateau. Their present-day altitudinal position is explained by passive ‘settling’ following disintegration of caprock and denudation of the underlying weaker rocks. Two localities are here presented in detail, Łężyckie Skałki and Pustelnik, along with adjacent boulder trains in the valleys incised into the plateau. It is argued that both localities have considerable geoheritage value and both play the role of geosites, although on-site facilities are so far limited. However, the complex history of boulders sets a series of challenges for successful geo-interpretation.

scholarly journals The Aspergillus nidulans Kinesin-3 UncA Motor Moves Vesicles along a Subpopulation of Microtubules

2009 ◽  
Vol 20 (2) ◽  
pp. 673-684 ◽  
Author(s):  
Nadine Zekert ◽  
Reinhard Fischer
Keyword(s):  
Aspergillus Nidulans ◽  
Filamentous Fungi ◽  
Fluorescent Protein ◽  
Hyphal Growth ◽  
Cellular Transport ◽  
Mitotic Spindles ◽  
Cytoplasmic Microtubules ◽  
Conventional Kinesin ◽  
Green Fluorescent ◽  
Vesicle Movement

The extremely polarized growth form of filamentous fungi imposes a huge challenge on the cellular transport machinery, because proteins and lipids required for hyphal extension need to be continuously transported to the growing tip. Recently, it was shown that endocytosis is also important for hyphal growth. Here, we found that the Aspergillus nidulans kinesin-3 motor protein UncA transports vesicles and is required for fast hyphal extension. Most surprisingly, UncA-dependent vesicle movement occurred along a subpopulation of microtubules. Green fluorescent protein (GFP)-labeled UncArigor decorated a single microtubule, which remained intact during mitosis, whereas other cytoplasmic microtubules were depolymerized. Mitotic spindles were not labeled with GFP-UncArigor but reacted with a specific antibody against tyrosinated α-tubulin. Hence, UncA binds preferentially to detyrosinated microtubules. In contrast, kinesin-1 (conventional kinesin) and kinesin-7 (KipA) did not show a preference for certain microtubules. This is the first example for different microtubule subpopulations in filamentous fungi and the first example for the preference of a kinesin-3 motor for detyrosinated microtubules.

scholarly journals Cut and paste – Efficient cotyledon micrografting for Arabidopsis thaliana seedlings

2019 ◽  
Author(s):  
Kai Bartusch ◽  
Jana Trenner ◽  
Marcel Quint
Keyword(s):  
Arabidopsis Thaliana ◽  
Success Rates ◽  
Long Distance ◽  
Long Distance Transport ◽  
Light Intensities ◽  
Whole Plant ◽  
Plant Level ◽  
Distance Transport ◽  
Plant Seedlings

AbstractCotyledon micrografting represents a very useful tool for studying the central role of cotyledons during early plant development, especially their interplay with other plant organs with regard to long distance transport. While hypocotyl grafting methods are established, cotyledon grafting is still inefficient. By optimizing cotyledon micrografting, we aim for higher success rates and increased throughput in the model species Arabidopsis thaliana. We established a cut and paste cotyledon surgery procedure on a flat solid but moist surface which improved handling of small plant seedlings. Applying a specific cutting and joining pattern throughput was increased up to 40 seedlings per hour. The combination of short day conditions and low light intensities for germination and long day plus high light intensities and vertical plate positioning after grafting significantly increased ‘ligation’ efficiency. Together, we achieved up to 46 % grafting success in A. thaliana. Reconnection of vasculature was shown by successful transport of a vasculature-specific dye across the grafting site. On a whole plant level, plants with grafted cotyledons match plants with intact cotyledons in biomass production and rosette development. This cut and paste cotyledon-to-petiole grafting protocol simplifies the handling of plant seedlings in surgery, increases the number of grafted plants per hour and produces higher success rates for A. thaliana seedlings. The developed cotyledon micrografting method is also suitable for other plant species of comparable size.

scholarly journals The role of roots and rhizosphere in providing tolerance to toxic metals and metalloids

2021 ◽  
Author(s):  
Dorina Podar ◽  
Frans J.M. Maathuis
Keyword(s):  
Negative Impact ◽  
Transport Processes ◽  
Long Distance ◽  
Root Cells ◽  
Metals And Metalloids ◽  
Natural Processes ◽  
Long Distance Transport ◽  
Bacteria And Fungi ◽  
Distance Transport

Human activity and natural processes have led to widespread dissemination of metals and metalloids, many of which are toxic and have a negative impact on agronomic production. Roots, as the first point of contact, are essential in endowing plants with tolerance to excess metal(loid) in the soil. The most important root responses include: adaptation of transport processes that affect uptake, efflux and long distance transport of metal(loid)s; metal(loid) detoxification within root cells via conjugation to thiol rich compounds and subsequent sequestration in the vacuole; plasticity in root architecture; the presence of bacteria and fungi in the rhizosphere that impact on metal(loid) bioavailability; the role of root exudates. In this review we will provide details on these processes and assess their relevance for the detoxification of arsenic, cadmium, mercury and zinc. Furthermore, we will assess if any of these methodologies has been tested in field conditions and whether they are effective in terms of improving crop metal(loid) tolerance.

scholarly journals The three isoenzymes of human inositol-1,4,5-trisphosphate 3-kinase show specific intracellular localization but comparable Ca2+ responses on transfection in COS-7 cells

2003 ◽  
Vol 374 (1) ◽  
pp. 41-49 ◽  
Author(s):  
Valérie DEWASTE ◽  
Colette MOREAU ◽  
Florence De SMEDT ◽  
Françoise BEX ◽  
Humbert De SMEDT ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Intracellular Localization ◽  
Metabolizing Enzymes ◽  
Transfected Cells ◽  
Extracellular Signals ◽  
Inositol 1,4,5 Trisphosphate ◽  
Low Levels ◽  
Green Fluorescent ◽  
Mutant Cells

Inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] 3-kinase catalyses the phosphorylation of InsP3 to inositol 1,3,4,5-tetrakisphosphate. cDNAs encoding three human isoenzymes of InsP3 3-kinase (A, B and C) have been reported previously [Choi, Kim, Lee, Moon, Sim, Kim, Chung and Rhee (1990) Science 248, 64–66; Dewaste, Pouillon, Moreau, Shears, Takazawa and Erneux (2000) Biochem. J. 352, 343–351; Dewaste, Roymans, Moreau and Erneux (2002) Biochem. Biophys. Res. Commun. 291, 400–405; Takazawa, Perret, Dumont and Erneux (1991) Biochem. Biophys. Res. Commun. 174, 529–535]. The localization of InsP3 3-kinase isoenzymes fused at their N-terminus to the green fluorescent protein has been studied by confocal microscopy. The A isoform appeared to associate with the cytoskeleton, whereas the C isoform was totally cytoplasmic. The B isoform had a more complex localization: it appeared in the plasma membrane, cytoskeleton and in the endoplasmic reticulum. The three human isoenzymes of InsP3 3-kinase can thus be distinguished by their N-terminal sequence, sensitivity to Ca2+/calmodulin and localization on transfection in COS-7 cells. We have compared the cytosolic Ca2+ responses induced by ATP in COS-7 cells transfected with the three isoenzymes. Cells expressing high levels of any of the three isoforms no longer respond to ATP, whereas cells expressing low levels of each enzyme showed a reduced response consisting of one to three Ca2+ spikes in response to 100 μM ATP. These effects were seen only in wild-type InsP3 3-kinase-transfected cells. 3-Kinase-dead mutant cells behaved as vector-transfected cells. The results highlight the potential role of the three isoforms of InsP3 3-kinase as direct InsP3 metabolizing enzymes and direct regulators of Ca2+ responses to extracellular signals.

scholarly journals Unravelling the involvement of cilevirus p32 protein in the viral transport

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mikhail Oliveira Leastro ◽  
Juliana Freitas-Astúa ◽  
Elliot Watanabe Kitajima ◽  
Vicente Pallás ◽  
Jesús A. Sánchez-Navarro
Keyword(s):  
Functional Study ◽  
Cell Periphery ◽  
Long Distance ◽  
Viral Spread ◽  
Long Distance Transport ◽  
Viral Transport ◽  
C Terminus ◽  
Citrus Leprosis ◽  
Distance Transport

AbstractCitrus leprosis (CL) is a severe disease that affects citrus orchards mainly in Latin America. It is caused by Brevipalpus-transmitted viruses from genera Cilevirus and Dichorhavirus. Currently, no reports have explored the movement machinery for the cilevirus. Here, we have performed a detailed functional study of the p32 movement protein (MP) of two cileviruses. Citrus leprosis-associated viruses are not able to move systemically in neither their natural nor experimental host plants. However, here we show that cilevirus MPs are able to allow the cell-to-cell and long-distance transport of movement-defective alfalfa mosaic virus (AMV). Several features related with the viral transport were explored, including: (i) the ability of cilevirus MPs to facilitate virus movement on a nucleocapsid assembly independent-manner; (ii) the generation of tubular structures from transient expression in protoplast; (iii) the capability of the N- and C- terminus of MP to interact with the cognate capsid protein (p29) and; (iv) the role of the C-terminus of p32 in the cell-to-cell and long-distance transport, tubule formation and the MP-plasmodesmata co-localization. The MP was able to direct the p29 to the plasmodesmata, whereby the C-terminus of MP is independently responsible to recruit the p29 to the cell periphery. Furthermore, we report that MP possess the capacity to enter the nucleolus and to bind to a major nucleolar protein, the fibrillarin. Based on our findings, we provide a model for the role of the p32 in the intra- and intercellular viral spread.

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