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51.
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The nucleotide substrate specificity of yeast poly(A) polymerase (yPAP) toward various C-2- and C-8-modified ATP analogs was examined. 32P-Radiolabeled RNA oligonucleotide primers were incubated with yPAP in the absence of ATP to assay polyadenylation using unnatural ATP substrates. The C-2-modified ATP analogs 2-amino-ATP and 2-chloro (Cl)-ATP were excellent substrates for yPAP. 8-Amino-ATP, 8-azido-ATP, and 8-aza-ATP all produced chain termination of polyadenylation, and no primer extension was observed with the C-8-halogenated derivatives 8-Br-ATP and 8-Cl-ATP. The effects of modified ATP analogs on ATP-dependent poly(A) tail synthesis by yPAP were also examined. Whereas C-2 substitution (2-amino-ATP and 2-Cl-ATP) had little effect on poly(A) tail length, C-8 substitution produced moderate (8-amino-ATP, 8-azido-ATP, and 8-aza-ATP) to substantial (8-Br-ATP and 8-Cl-ATP) reduction in poly(A) tail length. To model the biochemical consequences of 8-Cl-Ado incorporation into RNA primers, a synthetic RNA primer containing a 3'-terminal 8-Cl-AMP residue was prepared. Polyadenylation of this modified RNA primer by yPAP in the presence of ATP was blocked completely. To probe potential mechanisms of inhibition, two-dimensional NMR spectroscopy experiments were used to examine the conformation of two C-8-modified AMP nucleotides, 8-Cl-AMP and 8-amino-AMP. C-8 substitution in adenosine analogs shifted the ribose sugar pucker equilibrium to favor the DNA-like C-2'-endo form over the C-3'-endo (RNA-like) conformation, which suggests a potential mechanism for polyadenylation inhibition and chain termination. Base-modified ATP analogs may exert their biological effects through polyadenylation inhibition and thus may provide useful tools for investigating polyadenylation biochemistry within cells. 相似文献
53.
Gabbett TJ Sheppard JM Pritchard-Peschek KR Leveritt MD Aldred MJ 《Journal of strength and conditioning research / National Strength & Conditioning Association》2008,22(5):1413-1415
In this study, we evaluated the efficacy of two different dynamic warm-up conditions, one that was inclusive of open skills (i.e., reactive movements) and one that included only preplanned dynamic activities (i.e., closed skills) on the performance of speed, change of direction speed, vertical jump, and reactive agility in team sport athletes. Fourteen (six male, eight female) junior (mean +/- SD age, 16.3 +/- 0.7 year) basketball players participated in this study. Testing was conducted on 2 separate days using a within-subjects cross-over study design. Each athlete performed a standardized 7-minute warm-up consisting of general dynamic movements and stretching. After the general warm-up, athletes were randomly allocated into one of two groups that performed a dynamic 15-minute warm-up consisting entirely of open or closed skills. Each of the warm-up conditions consisted of five activities of 3 minute duration. At the completion of the warm-up protocol, players completed assessments of reactive agility, speed (5-, 10-, and 20-m sprints), change of direction speed (T-test), and vertical jump. No significant differences (p > 0.05) were detected among warm-up conditions for speed, vertical jump, change of direction speed, and reactive agility performances. The results of this study demonstrate that either open skill or closed skill warm-ups can be used effectively for team sport athletes without compromising performance on open skill and closed skill tasks. 相似文献
54.
Vlahakis NE Young BA Atakilit A Hawkridge AE Issaka RB Boudreau N Sheppard D 《The Journal of biological chemistry》2007,282(20):15187-15196
Vascular endothelial growth factor A (VEGF-A) is a potent inducer of angiogenesis. We now show that VEGF-A-induced adhesion and migration of human endothelial cells are dependent on the integrin alpha9beta1 and that VEGF-A is a direct ligand for this integrin. Adhesion and migration of these cells on the 165 and 121 isoforms of VEGF-A depend on cooperative input from alpha9beta1 and the cognate receptor for VEGF-A, VEGF receptor 2 (VEGF-R2). Unlike alpha3beta1or alphavbeta3 integrins, alpha9beta1 was also found to bind the 121 isoform of VEGF-A. This interaction appears to be biologically significant, because alpha9beta1-blocking antibody dramatically and specifically inhibited angiogenesis induced by VEGF-A165 or -121. Together with our previous findings that alpha9beta1 directly binds to VEGF-C and -D and contributes to lymphangiogenesis, these results identify the integrin alpha9beta1 as a potential pharmacotherapeutic target for inhibition of pathogenic angiogenesis and lymphangiogenesis. 相似文献
55.
Erdreich LS Van Kerkhove MD Scrafford CG Barraj L McNeely M Shum M Sheppard AR Kelsh M 《Radiation research》2007,168(2):253-261
Epidemiological studies of mobile phone use and risk of brain cancer have relied on self-reported use, years as a subscriber, and billing records as exposure surrogates without addressing the level of radiofrequency (RF) power output. The objective of this study was to measure environmental, behavioral and engineering factors affecting the RF power output of GSM mobile phones during operation. We estimated the RF-field exposure of volunteer subjects who made mobile phone calls using software-modified phones (SMPs) that recorded output power settings. Subjects recruited from three geographic areas in the U.S. were instructed to log information (place, time, etc.) for each call made and received during a 5-day period. The largest factor affecting energy output was study area, followed by user movement and location (inside or outside), use of a hands-free device, and urbanicity, although the two latter factors accounted for trivial parts of overall variance. Although some highly statistically significant differences were identified, the effects on average energy output rate were usually less than 50% and were generally comparable to the standard deviation. These results provide information applicable to improving the precision of exposure metrics for epidemiological studies of GSM mobile phones and may have broader application for other mobile phone systems and geographic locations. 相似文献
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Platelet-derived growth factor C (PDGF-C), a novel growth factor that binds to PDGF alpha and beta receptor 总被引:12,自引:0,他引:12
Gilbertson DG Duff ME West JW Kelly JD Sheppard PO Hofstrand PD Gao Z Shoemaker K Bukowski TR Moore M Feldhaus AL Humes JM Palmer TE Hart CE 《The Journal of biological chemistry》2001,276(29):27406-27414
We have characterized platelet-derived growth factor (PDGF) C, a novel growth factor belonging to the PDGF family. PDGF-C is a multidomain protein with the N-terminal region homologous to the extracellular CUB domain of neuropilin-1, and the C-terminal region consists of a growth factor domain (GFD) with homology to vascular endothelial growth factor (25%) and PDGF A-chain (23%). A serum-sensitive cleavage site between the two domains allows release of the GFD from the CUB domain. Competition binding and immunoprecipitation studies on cells bearing both PDGF alpha and beta receptors reveal a high affinity binding of recombinant GFD (PDGF-CC) to PDGF receptor-alpha homodimers and PDGF receptor-alpha/beta heterodimers. PDGF-CC exhibits greater mitogenic potency than PDGF-AA and comparable or greater mitogenic activity than PDGF-AB and PDGF-BB on several mesenchymal cell types. Analysis of PDGF-CC in vivo in a diabetic mouse model of delayed wound healing showed that PDGF-CC significantly enhanced repair of a full-thickness skin excision. Together, these studies describe a third member of the PDGF family (PDGF-C) as a potent mitogen for cells of mesenchymal origin in in vitro and in vivo systems with a binding pattern similar to PDGF-AB. 相似文献
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60.
Role of the cytoplasmic domain of the beta-subunit of integrin alpha(v)beta6 in infection by foot-and-mouth disease virus 总被引:1,自引:0,他引:1
Miller LC Blakemore W Sheppard D Atakilit A King AM Jackson T 《Journal of virology》2001,75(9):4158-4164
Field isolates of foot-and-mouth disease virus (FMDV) are believed to use RGD-dependent integrins as cellular receptors in vivo. Using SW480 cell transfectants, we have recently established that one such integrin, alpha(v)beta6, functions as a receptor for FMDV. This integrin was shown to function as a receptor for virus attachment. However, it was not known if the alpha(v)beta6 receptor itself participated in the events that follow virus binding to the host cell. In the present study, we investigated the effects of various deletion mutations in the beta6 cytoplasmic domain on infection. Our results show that although loss of the beta6 cytoplasmic domain has little effect on virus binding, this domain is essential for infection, indicating a critical role in postattachment events. The importance of endosomal acidification in alpha(v)beta6-mediated infection was confirmed by experiments showing that infection could be blocked by concanamycin A, a specific inhibitor of the vacuolar ATPase. 相似文献