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Cytoplasmic Components (cytoplasmic + component)
Selected AbstractsEstimation of variance components due to imprinting effects with DFREML and VCE: results of a simulation studyJOURNAL OF ANIMAL BREEDING AND GENETICS, Issue 1 2002A. ESSL Treating gametes as homozygous diploid individuals, TIER and SÖLKNER (Theor. Appl. Genet. 85: 868,872, 1993) proposed a method which manages the use of available computer programs with a common animal model to estimate variance components caused by imprinting effects. Despite some relevant model restrictions, this approach has already been used in some field data analyses by an adapted version of the widely used DFREML computer program, subsequently indicated by DFREMLa. The main objective of this study was to ascertain the properties of DFREMLa by computer simulation and to examine other alternative estimation approaches. The most important results may be summarized as follows: (1) Treating gametes as homozygous diploid individuals has the consequence that one-half of the actually realized gametic effect is totally abstracted in variance component estimation. Thus, an additional adjustment of the phenotypic variance calculated by DFREMLa is necessary to get correct values of estimated variance component ratios. (2) Adjusted DFREMLa estimates yielded correct results when animals were unselected and only maternal or paternal imprinting (not both simultaneously) occurred. (3) When the model did not adequately account for the additive genetic component within a maternal lineage, significant upward biases for the cytoplasmic component were observed. (4) The use of a simple dam and sire model with appropriate relationship matrices can be recommended when only the difference of maternal and paternal imprinting effects is of primary interest and the covariance between maternal halfsibs is not substantially increased by common environmental effects. (5) An adequate estimation of variance components for all possible imprinting situations requires the use of an animal model augmented by both maternal and paternal gametic effects. Unfortunately, a computer program on the basis of such a model does not yet exist. Schätzung von Varianzkomponenten für Imprintingeffekte mittels DFREML und VCE: Ergebnisse einer Simulationsstudie TIER and SÖLKNER (Theor. Appl. Genet. 85: 868,872, 1993) schlugen eine Methode zur Schätzung von imprintingbedingten Varianzkomponenten vor, die mit einem einfach zu adaptierenden Computerprogramm auf der Basis eines üblichen Tiermodells vorgenommen werden kann, indem sie Gameten wie homozygot diploide Individuen behandelten. Obwohl dieser Ansatz einige praxisrelevante Einschränkungen hat, wurde er bereits bei einigen Felddatenanalysen verwendet. Für diesen Zweck wurde eine entsprechend adaptierte Version des häufig verwendeten Computerprogrammes DFREML eingesetzt, die im folgenden mit DFREMLa bezeichnet wird. Das Ziel der vorliegen Untersuchung lag darin, die Eigenschaften von DFREMLa bei verschiedenen Imprintingsituationen zu überprüfen und weiters die Brauchbarkeit anderer möglicher Schätzansätze zu überprüfen. (1) Werden Gameten wie diploide homozygote Individuen aufgefaßt, dann geht bei der Schätzung von Varianzkomponenten mit DFREMLa eine Hälfte des tatsächlich wirksamen Gameteneffektes völlig verloren. Das heißt, die von DFREMLa ausgewiesenen Ergebnisse müssen nachträglich entsprechend adjustiert werden, um korrekte Schätzergebnisse für alle jene Quotienten von Varianzkomponenten zu erhalten, bei denen die gesamte phänotypische Varianz im Nenner steht. (2) Die adjustierten DFREMLa Schätzwerte lieferten in all jenen Fällen korrekte Ergebnisse, wo keine Selektion der Tiere erfolgte und entweder nur maternales oder paternales Imprinting (nicht beide gleichzeitig) auftrat. (3) Alle Modelle, bei denen die additiv genetische Komponente innerhalb einer Kuhfamilie keine adäquate Berücksichtigung fand, führten zu einer systematischen Überschätzung der zytoplasmatischen Varianzkomponente. (4) Ist nur jene Varianzkomponente von Interesse, die durch unterschiedlich starkes maternales bzw. paternales Imprinting erklärt werden kann, dann kann auch die Verwendung einfacher Vater-bzw. Muttermodelle empfohlen werden. Voraussetzung hierfür ist allerdings, daß die Kovarianz zwischen mütterlichen Halbgeschwistern durch keine gemeinsame Umwelt erhöht ist. (5) Eine für alle Imprintingsituationen problemadäquate Schätzung von Varianzkomponenten verlangt die Anwendung eines Tiermodelles, erweitert um beide imprintingbedingten Gameteneffekte. Leider fehlt gegenwärtig hierfür noch ein entsprechendes Computerprogramm. [source] Crystallization and preliminary X-ray analysis of FliJ, a cytoplasmic component of the flagellar type III protein-export apparatus from Salmonella sp.ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 1 2009Tatsuya Ibuki The axial component proteins of the bacterial flagellum are synthesized in the cytoplasm and then translocated into the central channel of the flagellum by the flagellar type III protein-export apparatus for self-assembly at the distal growing end of the flagellum. FliJ is an essential cytoplasmic component of the export apparatus. In this study, Salmonella FliJ with an extra three residues (glycine, serine and histidine) attached to the N-terminus as the remainder of a His tag (GSH-FliJ) was purified and crystallized. Crystals were obtained by the sitting-drop vapour-diffusion technique using PEG 300 as a precipitant. GSH-FliJ crystals grew in the hexagonal space group P6122 or P6522. While the native crystals diffracted to 3.3,Å resolution, the diffraction resolution limit of mercury derivatives was extended to 2.1,Å. Anomalous and isomorphous difference Patterson maps of the mercury-derivative crystal showed significant peaks in their Harker sections, indicating the usefulness of the derivative data for structure determination. [source] Cytoplasmic localization of oocyte-specific variant of porcine DNA methyltransferase-1 during early developmentDEVELOPMENTAL DYNAMICS, Issue 7 2009Young Sun Jeong Abstract DNA methyltransferase-1 (Dnmt1) is involved in the maintenance of genomic methylation patterns. Rather than full-length Dnmt1, mouse oocytes have a truncated variant called Dnmt1o. Immunofluorescence data showed that Dnmt1o localized to the cytoplasm, but this has not been confirmed using more direct methods. The cytoplasmic localization of Dnmt1o has been assigned to the main cause of global DNA demethylation in early mouse embryos. We studied localization of Dnmt1o in mouse and pig embryos. We identified pig Dnmt1o protein and its transcript with unique 5,-end sequence. Physically separating mouse and pig 2-cell embryos into their nuclear and cytoplasmic components demonstrated that Dnmt1o of both species localized to the cytoplasm. Cloned pig embryos had Dnmt1o as the main form, with no indication of somatic Dnmt1. These findings indicate that Dnmt1o is cytoplasmic during early development; its presence in both pig and mouse embryos further suggests that Dnmt1o is conserved in mammals. Developmental Dynamics 238:1666,1673, 2009. © 2009 Wiley-Liss, Inc. [source] Apparent mitochondrial asymmetry in Xenopus eggsDEVELOPMENTAL DYNAMICS, Issue 4 2003Natalia Volodina Abstract Cell polarity is manifest along the animal/vegetal axis in eggs of the frog, Xenopus laevis. Along this axis, maternal cytoplasmic components are asymmetrically distributed and are thought to underlie specification of distinct cell fates. To ascertain the molecular identities of such cytoplasmic components, we have used a monoclonal antibody that specifically stains the vegetal hemisphere of Xenopus eggs. The antigenic protein Vp67 (vegetal protein of 67 kDa) was identified through purification and cloning as a Xenopus homolog of the mitochondrial protein dihydrolipoamide acetyltransferase, a component of the pyruvate dehydrogenase complex. The identification of Vp67 as a mitochondrial protein could indicate that populations of mitochondria are asymmetrically distributed in Xenopus eggs. Developmental Dynamics 226:654,662, © 2003 Wiley-Liss, Inc. [source] The regulation of the endosomal compartment by p53 the tumor suppressor geneFEBS JOURNAL, Issue 8 2009Xin Yu The endosomal compartment of the cell is involved in a number of functions including: (a) internalizing membrane proteins to multivesicular bodies and lysosomes; (b) producing vesicles that are secreted from the cell (exosomes); and (c) generating autophagic vesicles that, especially in times of nutrient deprivation, supply cytoplasmic components to the lysosome for degradation and recycling of nutrients. The p53 protein responds to various stress signals by initiating a transcriptional program that restores cellular homeostasis and prevents the accumulation of errors in a cell. As part of this process, p53 regulates the transcription of a set of genes encoding proteins that populate the endosomal compartment and impact upon each of these endosomal functions. Here, we demonstrate that p53 regulates transcription of the genes TSAP6 and CHMP4C, which enhance exosome production, and CAV1 and CHMP4C, which produce a more rapid endosomal clearance of the epidermal growth factor receptor from the plasma membrane. Each of these p53-regulated endosomal functions results in the slowing of cell growth and division, the utilization of catabolic resources and cell-to-cell communication by exosomes after a stress signal is detected by the p53 protein. These processes avoid errors during stress and restore homeostasis once the stress is resolved. [source] Glucose-induced and nitrogen-starvation-induced peroxisome degradation are distinct processes in Hansenula polymorpha that involve both common and unique genesFEMS YEAST RESEARCH, Issue 1 2001Anna Rita Bellu Abstract In the methylotrophic yeast Hansenula polymorpha non-selective autophagy, induced by nitrogen starvation, results in the turnover of cytoplasmic components, including peroxisomes. We show that the uptake of these components occurs by invagination of the vacuolar membrane without their prior sequestration and thus differs from the mechanism described for bakers yeast. A selective mode of autophagy in H. polymorpha, namely glucose-induced peroxisome degradation, involves sequestration of individual peroxisomes tagged for degradation by membrane layers that subsequently fuse with the vacuole where the organelle is digested. H. polymorpha pdd mutants are blocked in selective peroxisome degradation. We observed that pdd1-201 is also impaired in non-selective autophagy, whereas this process still normally functions in pdd2-4. These findings suggest that mechanistically distinct processes as selective and non-selective autophagy involve common but also unique genes. [source] The crystal structure of microtubule-associated protein light chain 3, a mammalian homologue of Saccharomyces cerevisiae Atg8GENES TO CELLS, Issue 7 2004Kenji Sugawara Microtubule-associated protein light chain 3 (LC3), a mammalian homologue of yeast Atg8, plays an essential role in autophagy, which is involved in the bulk degradation of cytoplasmic components by the lysosomal system. Here, we report the crystal structure of LC3 at 2.05 Å resolution with an R-factor of 21.8% and a free R-factor of 24.9%. The structure of LC3, which is similar to those of Golgi-associated ATPase enhancer of 16 kDa (GATE-16) and GABAA receptor-associated protein (GABARAP), contains a ubiquitin core with two , helices, ,1 and ,2, attached at its N-terminus. Some common and distinct features are observed among these proteins, including the conservation of residues required to form an interaction among ,1, ,2 and the ubiquitin core. However, the electrostatic potential surfaces of these helices differ, implicating particular roles to select specific binding partners. Hydrophobic patches on the ubiquitin core of LC3, GABARAP and GATE-16 are well conserved and are similar to the E1 binding surface of ubiquitin and NEDD8. Therefore, we propose that the hydrophobic patch is a binding surface for mammalian Atg7 similar to a ubiquitin-like conjugation system. We also propose the functional implications of the ubiquitin fold as a recognition module of target proteins. [source] Modulation of nucleocytoplasmic trafficking by retention in cytoplasm or nucleusJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 6 2009Daniela M. Roth Abstract Nuclear protein transport processes have largely been studied using in vitro semi-intact cell systems where high concentrations of nuclear localizing substrates are used, and cytoplasmic components such as the microtubule (MT) network, are either absent or damaged. Here we use the fluorescence recovery after photobleaching (FRAP) technique to analyze the nucleocytoplasmic flux of distinct fluorescently tagged proteins over time in living cultured cells. FRAP was performed in different parts of the cell to analyze the kinetics of nucleocytoplasmic trafficking and intranuclear/cytoplasmic mobility of the tumor suppressor Rb protein and a SV40 large tumor antigen (T-ag) derivative containing the nuclear localization sequence (NLS), both fused to green fluorescent protein (GFP). The results indicate that proteins carrying the T-ag NLS are highly mobile in the nucleus and cytoplasm. Rb, in contrast, is largely immobile in both cellular compartments, with similar nuclear import and export kinetics. Rb nuclear export was CRM-1-mediated, with its reduced mobility in the cytoplasm in part due to association with MTs. Overall our results show that nuclear and cytoplasm retention modulates the rates of nuclear protein import and export in intact cells. J. Cell. Biochem. 107: 1160,1167, 2009. © 2009 Wiley-Liss, Inc. [source] SCANNING ELECTRON MICROSCOPY OBSERVATIONS OF DEFORMITIES IN SMALL PENNATE DIATOMS EXPOSED TO HIGH CADMIUM CONCENTRATIONS,JOURNAL OF PHYCOLOGY, Issue 6 2008Soizic Morin Different types of malformations are likely to affect the morphology of diatoms when exposed to particularly unstable environmental conditions, the most easily identifiable being distortion of the whole frustule. In the present study, we investigated, by means of SEM, valve abnormalities induced by high cadmium contamination (100 ,g · L,1) in small pennate diatoms. Changes in the shape of Amphora pediculus (Kütz.) Grunow and anomalous sculpturing of the cell wall of many species, such as Encyonema minutum (Hilse) D. G. Mann, Mayamaea agrestris (Hust.) Lange-Bert., Gomphonema parvulum (Kütz.) Kütz., or Eolimna minima (Grunow) Lange-Bert., were observed, which were not, or almost not, noticeable in the LM. With consideration to current knowledge of diatom morphogenesis, metal uptake by the cell would induce, directly or indirectly, damage to many cytoplasmic components (e.g., microtubules, cytoskeleton, Golgi-derived vesicles) involved in the precisely organized silica deposition. This study confirms that many species, whatever their size, are likely to exhibit morphological abnormalities under cadmium stress, and that this indicator may be valuable for the biomonitoring of metal contamination, even if SEM observations are not necessary for routine studies. [source] | |||||||||||||