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Pfluger SL Goodson HV Moran JM Ruggiero CJ Ye X Emmons KM Hager KM 《Eukaryotic cell》2005,4(2):432-442
Toxoplasma gondii and its apicomplexan relatives (such as Plasmodium falciparum, which causes malaria) are obligate intracellular parasites that rely on sequential protein release from specialized secretory organelles for invasion and multiplication within host cells. Because of the importance of these unusual membrane trafficking pathways for drug development and comparative cell biology, characterizing them is essential. In particular, it is unclear what role retrieval mechanisms play in parasite membrane trafficking or where they operate. Previously, we showed that T. gondii's beta-COP (TgBetaCOP; a subunit of coatomer protein complex I, COPI) and retrieval reporters localize exclusively to the zone between the parasite endoplasmic reticulum (ER) and Golgi apparatus. This suggested the existence of an HDEL receptor in T. gondii. We have now identified, cloned, and sequenced this receptor, TgERD2. TgERD2 localizes in a Golgi or ER pattern suggestive of the HDEL retrieval reporter (K. M. Hager, B. Striepen, L. G. Tilney, and D. S. Roos, J. Cell Sci. 112:2631-2638, 1999). A functional assay reveals that TgERD2 is able to complement the Saccharomyces cerevisiae ERD2 null mutant. Retrieval studies reveal that stable expression of a fluorescent exogenous retrieval ligand results in a dispersal of betaCOP signal throughout the cytoplasm and, surprisingly, results in betaCOP staining of the vacuolar space of the parasite. In contrast, stable expression of TgERD2GFP does not appear to disturb betaCOP staining. In addition to TgERD2, Toxoplasma contains two more divergent ERD2 relatives. Phylogenetic analysis reveals that these proteins belong to a previously unrecognized ERD2 subfamily common to plants and alveolate organisms and as such could represent mediators of parasite-specific retrieval functions. No evidence of class 2 ERD2 proteins was found in metazoan organisms or fungi. 相似文献
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Sang Jun Lee Paul T Pfluger Ji Young Kim Ruben Nogueiras Angeles Duran Gilles Pagès Jacques Pouysségur Matthias H Tschöp Maria T Diaz‐Meco Jorge Moscat 《EMBO reports》2010,11(3):226-232
In vivo genetic inactivation of the signalling adapter p62 leads to mature-onset obesity and insulin resistance, which correlate with reduced energy expenditure (EE) and increased adipogenesis, without alterations in feeding or locomotor functions. Enhanced extracellular signal-regulated kinase (ERK) activity in adipose tissue from p62-knockout (p62−/−) mice, and differentiating fibroblasts, suggested an important role for this kinase in the metabolic alterations of p62−/− mice. Here, we show that genetic inactivation of ERK1 in p62−/− mice reverses their increased adiposity and adipogenesis, lower EE and insulin resistance. These results establish genetically that p62 is a crucial regulator of ERK1 in metabolism. 相似文献
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Winker S.; Overbeek R.; Woese C.R.; OIsen G.J.; Pfluger N. 《Bioinformatics (Oxford, England)》1990,6(4):365-371
This paper summarizes our investigations into the computationaldetection of secondary and tertiary structure of ribosomal RNA.We have developed a new automated procedure that not only identifiespotential secondary and tertiary structural interactions, butalso provides the covariation evidence that supports the proposedbondings, and any counterevidence that can be detected in theknown sequences. A small number of previously unknown higher-orderstructural features have been detected in individual RNA molecules(16S rRNA and 7S RNA) through the use of our automated procedure.We are systematically studying mitochondrial rRNA, seeking tertiarystructure within 16S rRNA and quaternary structure between J6Sand23S rRNA. To test hypotheses suggested by an examination ofour program's output, our colleagues in biology are sequencingkey portions of the 23S ribosomal RNA for species in which theknown 16S ribosomal RNA exhibits variation (from the dominantpattern) at the site of a proposed bonding. Our ultimate hopeis that automated covariation analysis will contribute significantlyto a refined picture of ribosomal structure.
Received on January 17, 1990; accepted on June 1, 1990 相似文献
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Suzanne Arends Marianne Hofman Yvo PT Kamsma Eveline van der Veer Pieternella M Houtman Cees GM Kallenberg Anneke Spoorenberg Elisabeth Brouwer 《Arthritis research & therapy》2013,15(4):R99
Introduction
The aim of this study was to investigate the construct validity and test-retest reliability of the International Physical Activity Questionnaire (IPAQ; long form) and the Short QUestionnaire to Assess Health-enhancing physical activity (SQUASH) and to investigate the relation between daily physical activity and clinical assessments in patients with ankylosing spondylitis (AS).Methods
For validity, the self-report questionnaires IPAQ and SQUASH were compared with daily physical activity assessed with the ActiGraph accelerometer during 7 consecutive days in 63 AS outpatients. For reliability, the IPAQ and SQUASH were administered twice approximately 1 week apart in 52 AS outpatients. In all 115 patients, clinical assessments were performed at the outpatient clinic.Results
IPAQ and SQUASH total scores correlated significantly with accelerometer outcome: ρ = 0.38 and r = 0.35, respectively. Intraclass correlation coefficients between first and second assessments of the IPAQ and SQUASH were 0.83 and 0.89, respectively. Bland-Altman analyses showed no systemic bias, but in particular for the IPAQ the 95% limits of agreement were wide. Daily physical activity assessed by accelerometer, IPAQ, and SQUASH correlated significantly with disease activity, physical activity, and quality of life. A relation with spinal mobility was found only for the accelerometer and SQUASH. The direction of these correlations indicates that higher daily physical activity is related to lower disease activity and better physical function, spinal mobility and quality of life.Conclusions
Both physical activity questionnaires showed modest construct validity. The SQUASH showed good test-retest reliability, superior to the IPAQ. These results indicate that the SQUASH is more suitable than the IPAQ to assess daily physical activity in AS population studies. However, it is desirable to add questions on AS-specific physical activity. Further studies are needed to investigate the causality of the relation between daily physical activity and clinical assessments. 相似文献10.
Steven PT Hooton Andrew R Timms Joanna Rowsell Ray Wilson Ian F Connerton 《Virology journal》2011,8(1):1-14