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IRAG is essential for relaxation of receptor-triggered smooth muscle contraction by cGMP kinase
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Geiselhöringer A Werner M Sigl K Smital P Wörner R Acheo L Stieber J Weinmeister P Feil R Feil S Wegener J Hofmann F Schlossmann J 《The EMBO journal》2004,23(21):4222-4231
Signalling by cGMP-dependent protein kinase type I (cGKI) relaxes various smooth muscles modulating thereby vascular tone and gastrointestinal motility. cGKI-dependent relaxation is possibly mediated by phosphorylation of the inositol 1,4,5-trisphosphate receptor I (IP(3)RI)-associated protein (IRAG), which decreases hormone-induced IP(3)-dependent Ca(2+) release. We show now that the targeted deletion of exon 12 of IRAG coding for the N-terminus of the coiled-coil domain disrupted in vivo the IRAG-IP(3)RI interaction and resulted in hypomorphic IRAG(Delta12/Delta12) mice. These mice had a dilated gastrointestinal tract and a disturbed gastrointestinal motility. Carbachol- and phenylephrine-contracted smooth muscle strips from colon and aorta, respectively, of IRAG(Delta12/Delta12) mice were not relaxed by cGMP, while cAMP-mediated relaxation was unperturbed. Norepinephrine-induced increases in [Ca(2+)](i) were not decreased by cGMP in aortic smooth muscle cells from IRAG(Delta12/Delta12) mice. In contrast, cGMP-induced relaxation of potassium-induced smooth muscle contraction was not abolished in IRAG(Delta12/Delta12) mice. We conclude that cGMP-dependent relaxation of hormone receptor-triggered smooth muscle contraction essentially depends on the interaction of cGKI-IRAG with IP(3)RI. 相似文献
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Birgit Hoff Jens Kamerewerd Claudia Sigl Ivo Zadra Ulrich Kück 《Applied microbiology and biotechnology》2010,85(4):1081-1094
In Penicillium chrysogenum, the industrial producer of the β-lactam antibiotic penicillin, generating gene replacements for functional analyses is very
inefficient. Here, we constructed a recipient strain that allows efficient disruption of any target gene via homologous recombination.
Following isolation of the Pcku70 (syn. hdfA) gene encoding a conserved eukaryotic DNA-binding protein involved in non-homologous end joining (NHEJ), a Pcku70 knockout strain was constructed using a novel nourseothricin-resistance cassette as selectable marker. In detailed physiological
tests, strain ΔPcku70 showed no significant reduction in vegetative growth due to increased sensitivity to different mutagenic
substances. Importantly, deletion of the Pcku70 gene had no effect on penicillin biosynthesis. However, strain ΔPcku70 exhibits higher sensitivity to osmotic stress than
the parent strain. This correlated well with comparative data from microarray analyses: Genes related to the stress response
are significantly up-regulated in the Pcku70 deletion mutant. To demonstrate the applicability of strain ΔPcku70, three genes related to β-lactam antibiotic biosynthesis
were efficiently disrupted, indicating that this strain shows a low frequency of NHEJ, thus promoting efficient homologous
recombination. Furthermore, we discuss strategies to reactivate Pcku70 in strains successfully used for gene disruptions. 相似文献
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Tomek-Roksandić S Tomasović-Mrcela N Narancić NS Sigl G 《Collegium antropologicum》2010,34(3):841-846
Gerontology-public health indicators of functional ability of the elderly in institutional and non-institutional health care in Croatia were determined by use of expert methodology developed at Department of Gerontology, Dr. Andrija Stampar Institute of Public Health in Zagreb, with the aim to upgrade the Program of Health Care Measures and Procedures in Health Care of the Elderly. Comparison of functional ability between the users of selected Old People's Homes (institutional care; N = 5030) and Gerontology Centers (non-institutional care; N = 2112) yielded highest between-group difference in the proportion of "fully movable" and "fully independent" categories in favor of the latter, thus steering the program of health care for the elderly accordingly. In addition, study results showed greater difference in the proportion of categories describing mental status of institutional and non-institutional care users as compared with the categories describing their physical status, suggesting that mental status plays a more important role than physical status in the geriatric user's stay in non-institutional care versus institutional care. This issue requires additional studies. The results obtained by this indicator analysis pointed to the preventive and geroprophylactic measures to ensure efficient health care for the elderly and to develop the program of mental health promotion and preservation. According to 2007 estimate, there were 759,318 (16.9%) persons aged > or = 65 in Croatia. Data collected at gerontology database kept at Department of Gerontology, Dr. Andrija Stampar Institute of Public Health (September 2008) showed 2% of the elderly (N = 14807) to be accommodated at Old People's Homes, which is below the European average of 4%. 相似文献
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A Steidle K Sigl R Schuhegger A Ihring M Schmid S Gantner M Stoffels K Riedel M Givskov A Hartmann C Langebartels L Eberl 《Applied and environmental microbiology》2001,67(12):5761-5770
Given that a large proportion of the bacteria colonizing the roots of plants is capable of producing N-acyl-L-homoserine lactone (AHL) molecules, it appears likely that these bacterial pheromones may serve as signals for communication between cells of different species. In this study, we have developed and characterized novel Gfp-based monitor strains that allow in situ visualization of AHL-mediated communication between individual cells in the plant rhizosphere. For this purpose, three Gfp-based AHL sensor plasmids that respond to different spectra of AHL molecules were transferred into AHL-negative derivatives of Pseudomonas putida IsoF and Serratia liquefaciens MG1, two strains that are capable of colonizing tomato roots. These AHL monitor strains were used to visualize communication between defined bacterial populations in the rhizosphere of axenically grown tomato plants. Furthermore, we integrated into the chromosome of AHL-negative P. putida strain F117 an AHL sensor cassette that responds to the presence of long-chain AHLs with the expression of Gfp. This monitor strain was used to demonstrate that the indigenous bacterial community colonizing the roots of tomato plants growing in nonsterile soil produces AHL molecules. The results strongly support the view that AHL signal molecules serve as a universal language for communication between the different bacterial populations of the rhizosphere consortium. 相似文献
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Sigl H Brink G Seufert M Schulz M Wegner G Sackmann E 《European biophysics journal : EBJ》1997,25(4):249-259
The present work deals with the assembly of multilayers or rod-like polymers with hydrophobic side chains (called hairy rods)
and their potential application as ultrathin polymer cushions for the build-up of self healing supported membranes on various
solids (Si/SiO2-wafer, gold covered substrates). Three types of hairy rods were studied: Isopentyl cellulose (IPC), phtalocyaniatopolysiloxane
with mixed alkane side chains (PCPS) and trimethylsilane cellulose (TMCS). Detailed analysis of the thickness of supported
multilayers as a function of the number of deposited monolayers with ellipsometry, near infrared surface plasmon resonance
(NIR-SPR), a quartz crystal microbalance (QCM) and reflection interference contrast microscopy (RICM), show that the basic
building blocks of hairy rod multilayers are bilayers with the hydrophobic surfaces of the monolayers facing each other. Continuous
and stable firms of hairy rods can be deposited if the hydrophobicities of the solid surface and the monolayer are matched.
It is demonstrated by lateral diffusion measurements (using photobleaching techniques) that continuous phospholipid bilayers
can be deposited onto multilayers of rigid rods of TMCS after hydrophilization by cleavage of trimethylsilane side chains
in HCl-vapour, while stable lipid monolayers can be deposited onto hydrophobic surfaces of rigid rod layers. NIR-SPR allows
the observation of double band reflectivity curves at interfaces separating different surface layers and thus offers the possibility
of differential detection of ligand binding at the interface of differently functionalized domains.
Received: 2 February 1996 / Accepted: 28 October 1996 相似文献
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Reinhard Sigl Christian Ploner Giridhar Shivalingaiah Reinhard Kofler Stephan Geley 《PloS one》2014,9(5)
RNA interference (RNAi) has become an essential technology for functional gene analysis. Its success, however, depends on the effective expression of RNAi-inducing small double-stranded interfering RNA molecules (siRNAs) in target cells. In many cell types, RNAi can be achieved by transfection of chemically synthesised siRNAs, which results in transient knockdown of protein expression. Expression of double-stranded short hairpin RNA (shRNA) provides another means to induce RNAi in cells that are hard to transfect. To facilitate the generation of stable, conditional RNAi cell lines, we have developed novel one- and two-component vector GATEWAY-compatible lentiviral tetracycline-regulated RNAi (GLTR) systems. The combination of a modified RNA-polymerase-III-dependent H1 RNA promoter (designated ‘THT’) for conditional shRNA expression with different lentiviral delivery vectors allows (1) the use of fluorescent proteins for colour-coded combinatorial RNAi or for monitoring RNAi induction (pGLTR-FP), (2) selection of transduced cells (pGLTR-S), and (3) the generation of conditional cell lines using a one vector system (pGLTR-X). All three systems were found to be suitable for the analysis of essential genes, such as CDC27, a component of the mitotic ubiquitin ligase APC/C, in cell lines and primary human cells. 相似文献
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Sebastian Steibl Robert Sigl Sanja Blaha Sophia Drescher Gerhard Gebauer Elif Gürkal Frederic Hüftlein Anna Satzger Michael Schwarzer Dimitri Seidenath Jana Welfenbach Raphael S. Zinser Christian Laforsch 《Ecology and evolution》2021,11(19):13128
Ecosystems are interconnected by energy fluxes that provide resources for the inhabiting organisms along the transition zone. Especially where in situ resources are scarce, ecosystems can become highly dependent on external resources. The dependency on external input becomes less pronounced in systems with elevated in situ production, where only consumer species close to the site of external input remain subsidized, whereas species distant to the input site rely on the in situ production of the ecosystem. It is largely unclear though if this pattern is consistent over different consumer species and trophic levels in one ecosystem, and whether consumer species that occur both proximate to and at a distance from the input site differ in their dependency on external resource inputs between sites. Using stable isotope analysis, we investigated the dependency on external marine input for common ground‐associated consumer taxa on small tropical islands with high in situ production. We show that marine input is only relevant for strict beach‐dwelling taxa, while the terrestrial vegetation is the main carbon source for inland‐dwelling taxa. Consumer species that occurred both close (beach) and distant (inland) to the site of marine input showed similar proportions of marine input in their diets. This supports earlier findings that the relevance of external resources becomes limited to species close to the input site in systems with sufficient in situ production. However, it also indicates that the relevance of external input is also species‐dependent, as consumers occurring close and distant to the input site depended equally strong or weak on marine input. 相似文献
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C Wandke M Barisic R Sigl V Rauch F Wolf AC Amaro CH Tan AJ Pereira U Kutay H Maiato P Meraldi S Geley 《The Journal of cell biology》2012,198(5):847-863
Chromokinesins are microtubule plus end-directed motor proteins that bind to chromosome arms. In Xenopus egg cell-free extracts, Xkid and Xklp1 are essential for bipolar spindle formation but the functions of the human homologues, hKID (KIF22) and KIF4A, are poorly understood. By using RNAi-mediated protein knockdown in human cells, we find that only co-depletion delayed progression through mitosis in a Mad2-dependent manner. Depletion of hKID caused abnormal chromosome arm orientation, delayed chromosome congression, and sensitized cells to nocodazole. Knockdown of KIF4A increased the number and length of microtubules, altered kinetochore oscillations, and decreased kinetochore microtubule flux. These changes were associated with failures in establishing a tight metaphase plate and an increase in anaphase lagging chromosomes. Co-depletion of both chromokinesins aggravated chromosome attachment failures, which led to mitotic arrest. Thus, hKID and KIF4A contribute independently to the rapid and correct attachment of chromosomes by controlling the positioning of chromosome arms and the dynamics of microtubules, respectively. 相似文献