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31.
Sharma M Leung L Brocardo M Henderson J Flegg C Henderson BR 《The Journal of biological chemistry》2006,281(25):17140-17149
Adenomatous polyposis coli protein (APC) translocates to, and stabilizes, the plus-ends of microtubules. In microtubule-dependent cellular protrusions, APC frequently accumulates in peripheral clusters at the basal membrane. APC targeting to membrane clusters is important for cell migration, but the localization mechanism is poorly understood. In this study, we performed deletion mapping and defined a minimal sequence (amino acids 1-2226) that efficiently targets APC to membrane clusters. This sequence lacks DLG-1 and EB1 binding sites, suggesting that these partners are not absolutely required for APC membrane targeting. A series of APC sequences were transiently expressed in cells and compared for their ability to compete endogenous APC at the membrane; potent inhibition of endogenous APC targeting was elicited by the Armadillo- (binds KAP3A, B56alpha, and ASEF) and beta-catenin-binding domains. The Armadillo domain was predicted to inhibit APC membrane localization through sequestration of the kinesin-KAP3A complex. The role of beta-catenin in APC membrane localization was unexpected but affirmed by overexpressing the APC binding sequence of beta-catenin, which similarly reduced APC membrane staining. Furthermore, we used RNA interference to show that loss of beta-catenin reduced APC at membrane clusters in migrating cells. In addition, we report that transiently expressed APC-yellow fluorescent protein co-localized with beta-catenin, KAP3A, EB1, and DLG-1 at membrane clusters, but only beta-catenin stimulated APC anchorage at the membrane. Our findings identify beta-catenin as a regulator of APC targeting to membrane clusters and link these two proteins to cell migration. 相似文献
32.
The length but not the sequence of the polyoma virus late leader exon is important for both late RNA splicing and stability. 总被引:10,自引:2,他引:10 下载免费PDF全文
Polyoma virus late RNA processing provides a convenient model system in which to study the mechanics of splicing in vivo. In order to understand further the role of the untranslated "late leader" unit in late RNA processing we have constructed a group of polyoma viruses with deletions and substitutions in the leader exon. This has allowed us to determine that there is a minimum exon size required for both pre-mRNA splicing and stability in this system. We show here that the non-viability of a mutant (ALM) with a 9 base late leader unit is due to a general defect in late RNA splicing. In addition, ALM-infected cells show at least 40-fold depression in the accumulation of late nuclear RNA (spliced or unspliced). The ALM late promoter, however, functions nearly normally. Substituted leader variants with 51- to 96-base long exons of unrelated sequence are viable (G. Adami and G. Carmichael, J. Virol. 58, 417-425, 1986). We show here that late RNA from one of these substituted leader mutants (containing a 51-base leader exon) is spliced at wild type levels, with virtually no defect in accumulation. Thus, in the polyoma system, splice sites separated by only 9 bases can inhibit each others usage, presumably by steric interference. We suggest that this type of inhibition leads to extreme RNA instability. 相似文献
33.
Evidence for a dinuclear active site in the metallo-beta-lactamase BcII with substoichiometric Co(II). A new model for metal uptake 总被引:1,自引:0,他引:1
Llarrull LI Tioni MF Kowalski J Bennett B Vila AJ 《The Journal of biological chemistry》2007,282(42):30586-30595
Metallo-beta-lactamases are zinc-dependent enzymes that constitute one of the main resistance mechanisms to beta-lactam antibiotics. Metallo-beta-lactamases have been characterized both in mono- and dimetallic forms. Despite many studies, the role of each metal binding site in substrate binding and catalysis is still unclear. This is mostly due to the difficulties in assessing the metal content and site occupancy in solution. For this reason, Co(II) has been utilized as a useful probe of the active site structure. We have employed UV-visible, EPR, and NMR spectroscopy to study Co(II) binding to the metallo-beta-lactamase BcII from Bacillus cereus. The spectroscopic features were attributed to the two canonical metal binding sites, the 3H (His(116), His(118), and His(196)) and DCH (Asp(120), Cys(221), and His(263)) sites. These data clearly reveal the coexistence of mononuclear and dinuclear Co(II)-loaded forms at Co(II)/enzyme ratios as low as 0.6. This picture is consistent with the macroscopic dissociation constants here determined from competition binding experiments. A spectral feature previously assigned to the DCH site in the dinuclear species corresponds to a third, weakly bound Co(II) site. The present work emphasizes the importance of using different spectroscopic techniques to follow the metal content and localization during metallo-beta-lactamase turnover. 相似文献
34.
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36.
María Sol Gaspe Yael Mariana Provecho Romina Valeria Piccinali Ricardo Esteban Gürtler/ 《Memórias do Instituto Oswaldo Cruz》2015,110(3):310-318
House re-invasion by native triatomines after insecticide-based control campaigns
represents a major threat for Chagas disease vector control. We conducted a
longitudinal intervention study in a rural section (Area III, 407 houses) of Pampa
del Indio, northeastern Argentina, and used wing geometric morphometry to compare
pre-spray and post-spray (re-infestant bugs) Triatoma infestans
populations. The community-wide spraying with pyrethroids reduced the prevalence of
house infestation by T. infestans from 31.9% to < 1% during a
four-year follow-up, unlike our previous studies in the neighbouring Area I. Two
groups of bug collection sites differing in wing shape variables before interventions
(including 221 adults from 11 domiciles) were used as a reference for assigning 44
post-spray adults. Wing shape variables from post-spray, high-density bug colonies
and pre-spray groups were significantly different, suggesting that re-infestant
insects had an external origin. Insects from one house differed strongly in wing
shape variables from all other specimens. A further comparison between insects from
both areas supported the existence of independent re-infestation processes within the
same district. These results point to local heterogeneities in house re-infestation
dynamics and emphasise the need to expand the geographic coverage of vector
surveillance and control operations to the affected region. 相似文献
37.
Mark T. Gladwin Robyn J. Barst J. Simon R. Gibbs Mariana Hildesheim Vandana Sachdev Mehdi Nouraie Kathryn L. Hassell Jane A. Little Dean E. Schraufnagel Lakshmanan Krishnamurti Enrico Novelli Reda E. Girgis Claudia R. Morris Erika Berman Rosenzweig David B. Badesch Sophie Lanzkron Oswaldo L. Castro James G. Taylor VI Jonathan C. Goldsmith Gregory J. Kato Victor R. Gordeuk Roberto F. Machado 《PloS one》2014,9(7)
Background
The role of pulmonary hypertension as a cause of mortality in sickle cell disease (SCD) is controversial.Methods and Results
We evaluated the relationship between an elevated estimated pulmonary artery systolic pressure and mortality in patients with SCD. We followed patients from the walk-PHaSST screening cohort for a median of 29 months. A tricuspid regurgitation velocity (TRV)≥3.0 m/s cuttof, which has a 67–75% positive predictive value for mean pulmonary artery pressure ≥25 mm Hg was used. Among 572 subjects, 11.2% had TRV≥3.0 m/sec. Among 582 with a measured NT-proBNP, 24.1% had values ≥160 pg/mL. Of 22 deaths during follow-up, 50% had a TRV≥3.0 m/sec. At 24 months the cumulative survival was 83% with TRV≥3.0 m/sec and 98% with TRV<3.0 m/sec (p<0.0001). The hazard ratios for death were 11.1 (95% CI 4.1–30.1; p<0.0001) for TRV≥3.0 m/sec, 4.6 (1.8–11.3; p = 0.001) for NT-proBNP≥160 pg/mL, and 14.9 (5.5–39.9; p<0.0001) for both TRV≥3.0 m/sec and NT-proBNP≥160 pg/mL. Age >47 years, male gender, chronic transfusions, WHO class III–IV, increased hemolytic markers, ferritin and creatinine were also associated with increased risk of death.Conclusions
A TRV≥3.0 m/sec occurs in approximately 10% of individuals and has the highest risk for death of any measured variable.The study is registered in ClinicalTrials.gov with identifier
NCT00492531相似文献38.
Mariana Vlad E. Bordas Rodica Tomus Doina Sava Ecaterina Farkas G. Uza 《Biological trace element research》1993,38(1):47-54
Serum copper concentration increases significantly (p<0.01) in rats with experimental atherosclerosis compared to a control group. The serum zinc, the zinc, and copper concentration
in abdominal aorta and in liver decreases significantly (p<0.05) compared to the control group. Administration of copper sulfate for 100 d in these animals induces a significant increase
of serum copper (p<0.01), decrease of serum cholesterol (p<0.05) and increase of liver copper concentration as compared with the group fed only a high cholesterol diet. In the aorta
of these animals the copper concentration increases and edema and lipid infiltration are considerably less than in the group
of animals fed only a high lipid diet. 相似文献
39.
Photodynamic inactivation is a new promising approach to treat bacterial infections. Usually, the evaluation of the efficacy
of this method is done through time-consuming and labor-intensive microbiological test methods. This paper describes the development
and implementation of an optical method to evaluate the photodynamic inactivation of bacteria based on non-invasive diffuse
reflectance measurements. Five Staphylococcus aureus cultures and 15 mice have been used in this study. A skin lesion was created on the back of all animals, and it was contaminated
with S. aureus (5.16 ± 0.013 log CFU/ml). Toluidine Blue O (c = 8.67 × 10 − 3 M) has been used as a photosensitiser agent. The bacterial cultures and animals were exposed to laser radiation (λ = 635 nm, P = 15 mW, DE = 8.654 J/cm2) for 20 min. The photodynamic inactivation of bacteria was monitored by acquiring the wounds’ reflection spectra at different
time points and by microbiological exams on the bioptical material. The good correlation between the diffuse reflectance and
colony-forming units demonstrates the value of this optical method based on diffuse reflectance measurements as a rapid technique
to monitor photodynamic bacterial inactivation. 相似文献
40.
Silva-Brandão KL Lyra ML Santos TV Seraphim N Albernaz KC Pavinato VA Martinelli S Cônsoli FL Omoto C 《Genetics and molecular biology》2011,34(4):719-725
The applicability of mitochondrial nad6 sequences to studies of DNA and population variability in Lepidoptera was tested in four species of economically important moths and one of wild butterflies. The genetic information so obtained was compared to that of cox1 sequences for two species of Lepidoptera. nad6 primers appropriately amplified all the tested DNA targets, the generated data proving to be as informative and suitable in recovering population structures as that of cox1. The proposal is that, to obtain more robust results, this mitochondrial region can be complementarily used with other molecular sequences in studies of low level phylogeny and population genetics in Lepidoptera. 相似文献