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191.
192.
Raney T 《Current opinion in biotechnology》2006,17(2):174-178
Transgenic crops are being adopted rapidly at the global level, but only a few developing countries are growing them in significant quantities. Why are these crops so successful in some countries but not in others? Farm level profitability ultimately determines whether farmers adopt and retain a new technology, but this depends on much more than technical performance. Recent economic studies in developing countries find positive, but highly variable, economic returns to adopting transgenic crops. These studies confirm that institutional factors such as national agricultural research capacity, environmental and food safety regulations, intellectual property rights and agricultural input markets matter at least as much as the technology itself in determining the level and distribution of economic benefits. 相似文献
193.
Murray JM O'Neill JP Messier T Rivers J Walker VE McGonagle B Trombley L Cowell LG Kelsoe G McBlane F Finette BA 《Journal of immunology (Baltimore, Md. : 1950)》2006,177(8):5393-5404
V(D)J recombinase mediates rearrangements at immune loci and cryptic recombination signal sequences (cRSS), resulting in a variety of genomic rearrangements in normal lymphocytes and leukemic cells from children and adults. The frequency at which these rearrangements occur and their potential pathologic consequences are developmentally dependent. To gain insight into V(D)J recombinase-mediated events during human development, we investigated 265 coding junctions associated with cRSS sites at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) locus in peripheral T cells from 111 children during the late stages of fetal development through early adolescence. We observed a number of specific V(D)J recombinase processing features that were both age and gender dependent. In particular, TdT-mediated nucleotide insertions varied depending on age and gender, including percentage of coding junctions containing N-nucleotide inserts, predominance of GC nucleotides, and presence of inverted repeats (Pr-nucleotides) at processed coding ends. In addition, the extent of exonucleolytic processing of coding ends was inversely related to age. We also observed a coding-partner-dependent difference in exonucleolytic processing and an age-specific difference in the subtypes of V(D)J-mediated events. We investigated these age- and gender-specific differences with recombination signal information content analysis of the cRSS sites in the human HPRT locus to gain insight into the mechanisms mediating these developmentally specific V(D)J recombinase-mediated rearrangements in humans. 相似文献
194.
Yvette R. Pittman Kimberly Kandl Marcus Lewis Louis Valente Terri Goss Kinzy 《The Journal of biological chemistry》2009,284(7):4739-4747
Eukaryotic translation elongation factor 1A (eEF1A) both shuttles
aminoacyl-tRNA (aa-tRNA) to the ribosome and binds and bundles actin. A single
domain of eEF1A is proposed to bind actin, aa-tRNA and the guanine nucleotide
exchange factor eEF1Bα. We show that eEF1Bα has the ability to
disrupt eEF1A-induced actin organization. Mutational analysis of eEF1Bα
F163, which binds in this domain, demonstrates effects on growth, eEF1A
binding, nucleotide exchange activity, and cell morphology. These phenotypes
can be partially restored by an intragenic W130A mutation. Furthermore, the
combination of F163A with the lethal K205A mutation restores viability by
drastically reducing eEF1Bα affinity for eEF1A. This also results in a
consistent increase in actin bundling and partially corrected morphology. The
consequences of the overlapping functions in this eEF1A domain and its unique
differences from the bacterial homologs provide a novel function for
eEF1Bα to balance the dual roles in actin bundling and protein
synthesis.The final step of gene expression takes place at the ribosome as mRNA is
translated into protein. In the yeast Saccharomyces cerevisiae,
elongation of the polypeptide chain requires the orchestrated action of three
soluble factors. The eukaryotic elongation factor 1
(eEF1)2 complex
delivers aminoacyl-tRNA (aa-tRNA) to the empty A-site of the elongating
ribosome (1). The eEF1A subunit
is a classic G-protein that acts as a “molecular switch” for the
active and inactive states based on whether GTP or GDP is bound, respectively
(2). Once an anticodon-codon
match occurs, the ribosome acts as a GTPase-activating factor to stimulate GTP
hydrolysis resulting in the release of inactive GDP-bound eEF1A from the
ribosome. Because the intrinsic rate of GDP release from eEF1A is extremely
slow (3,
4), a guanine nucleotide
exchange factor (GEF) complex, eEF1B, is required
(5,
6). The yeast S.
cerevisiae eEF1B complex contains two subunits, the essential catalytic
subunit eEF1Bα (5) and
the non-essential subunit eEF1Bγ
(7).The co-crystal structures of eEF1A:eEF1Bα C terminus:GDP:
Mg2+ and eEF1A:eEF1Bα C terminus:GDPNP
(8,
9) demonstrated a surprising
structural divergence from the bacterial EF-Tu-EF-Ts
(10) and mammalian
mitochondrial EF-Tumt-EF-Tsmt
(11). While the G-proteins
have a similar topology and consist of three well-defined domains, a striking
difference was observed in binding sites for their GEFs. The C terminus of
eEF1Bα interacts with domain I and a distinct pocket of domain II eEF1A,
creating two binding interfaces. In contrast, the bacterial counterpart EF-Ts
and mammalian mitochondrial EF-Tsmt, make extensive contacts with
domain I and III of EF-Tu and EF-Tumt, respectively. The altered
binding interface of eEF1Bα to domain II of eEF1A is particularly
unexpected given the functions associated with domain II of eEF1A and EF-Tu.
The crystal structure of the EF-Tu:GDPNP:Phe-tRNAPhe complex
reveals aa-tRNA binding to EF-Tu requires only minor parts of both domain II
and tRNA to sustain stable contacts
(12). That eEF1A employs the
same aa-tRNA binding site is supported by genetic and biochemical data
(13-15).
Interestingly, eEF1Bα contacts many domain II eEF1A residues in the
region hypothesized to be involved in the binding of the aa-tRNA CCA end
(8). Because, the shared
binding site of eEF1Bα and aa-tRNA on domain II of eEF1A is
significantly different between the eukaryotic and bacterial/mitochondrial
systems, eEF1Bα may play a unique function aside from guanine nucleotide
release in eukaryotes.In eukaroytes, eEF1A is also an actin-binding and -bundling protein. This
noncanonical function of eEF1A was initially observed in Dictyostelium
amoebae (16). It is
estimated that greater than 60% of Dictyostelium eEF1A is associated
with the actin cytoskeleton
(17). The eEF1A-actin
interaction is conserved among species from yeast to mammals, suggesting the
importance of eEF1A for cytoskeleton integrity. Using a unique genetic
approach, multiple eEF1A mutations were identified that altered cell growth
and morphology, and are deficient in bundling actin in vitro
(18,
19). Intriguingly, most
mutations localized to domain II, the shared aa-tRNA and eEF1Bα binding
site. Previous studies have demonstrated that actin bundling by eEF1A is
significantly reduced in the presence of aa-tRNA while eEF1A bound to actin
filaments is not in complex with aa-tRNA
(20). Therefore, actin and
aa-tRNA binding to eEF1A is mutually exclusive. In addition, overexpression of
yeast eEF1A or actin-bundling deficient mutants do not affect translation
elongation (18,
19,
21), suggesting
eEF1A-dependent cytoskeletal organization is independent of its translation
elongation function (18,
20). Thus, while aa-tRNA
binding to domain II is conserved between EF-Tu and eEF1A, this actin bundling
function associated with eEF1A domain II places greater importance on its
relationship with the “novel” binding interface between eEF1A
domain II and eEF1Bα.Based on this support for an overlapping actin bundling and eEF1Bα
binding site in eEF1A domain II, we hypothesize that eEF1Bα modulates
the equilibrium between actin and translation functions of eEF1A and is
perhaps the result of evolutionary selective pressure to balance the
eukaryotic-specific role of eEF1A in actin organization. Here, we present
kinetic and biochemical evidence using a F163A mutant of eEF1Bα for the
importance of the interactions between domain II of eEF1A and eEF1Bα to
prevent eEF1A-dependent actin bundling as well as promoting guanine nucleotide
exchange. Furthermore, altered affinities of eEF1Bα mutants for eEF1A
support that this complex formation is a determining factor for eEF1A-induced
actin organization. Interestingly, the F163A that reduces eEF1A affinity is an
intragenic suppressor of the lethal K205A eEF1Bα mutant that displays
increased affinity for eEF1A. This, along with a consistent change in the
actin bundling correlated with the affinity of eEF1Bα for eEF1A,
indicates that eEF1Bα is a balancer, directing eEF1A to translation
elongation and away from actin, and alterations in this balance result in
detrimental effects on cell growth and eEF1A function. 相似文献
195.
Torgerson DG Capurso D Mathias RA Graves PE Hernandez RD Beaty TH Bleecker ER Raby BA Meyers DA Barnes KC Weiss ST Martinez FD Nicolae DL Ober C 《American journal of human genetics》2012,90(2):273-281
Common variation in over 100 genes has been implicated in the risk of developing asthma, but the contribution of rare variants to asthma susceptibility remains largely unexplored. We selected nine genes that showed the strongest signatures of weak purifying selection from among 53 candidate asthma-associated genes, and we sequenced the coding exons and flanking noncoding regions in 450 asthmatic cases and 515 nonasthmatic controls. We observed an overall excess of p values <0.05 (p = 0.02), and rare variants in four genes (AGT, DPP10, IKBKAP, and IL12RB1) contributed to asthma susceptibility among African Americans. Rare variants in IL12RB1 were also associated with asthma susceptibility among European Americans, despite the fact that the majority of rare variants in IL12RB1 were specific to either one of the populations. The combined evidence of association with rare noncoding variants in IL12RB1 remained significant (p = 3.7 × 10(-4)) after correcting for multiple testing. Overall, the contribution of rare variants to asthma susceptibility was predominantly due to noncoding variants in sequences flanking the exons, although nonsynonymous rare variants in DPP10 and in IL12RB1 were associated with asthma in African Americans and European Americans, respectively. This study provides evidence that rare variants contribute to asthma susceptibility. Additional studies are required for testing whether prioritizing genes for resequencing on the basis of signatures of purifying selection is an efficient means of identifying novel rare variants that contribute to complex disease. 相似文献
196.
Amy R. Kireta Euan D. Reavie Gerald V. Sgro Ted R. Angradi David W. Bolgrien Terri M. Jicha Brian H. Hill 《Hydrobiologia》2012,691(1):171-188
Diatom-based indicators were developed to assess environmental conditions in the Missouri, Ohio, and Upper Mississippi rivers. Disturbance gradients, comprising the first two principal components derived from a suite of stressor variables, included a trophic gradient (Trophic) and a gradient reflecting agriculture and other development activities (Ag/Dev). Diatom-based indicators were developed by creating models using weighted average calibration and regression-based transfer functions to relate planktonic and periphytic diatom species assemblages to each disturbance gradient. The most predictive disturbance models combined phytoplankton and periphyton assemblages into a single bioindicator model (observed versus inferred: Trophic $ r_{\text{boot}}^{2} = 0. 5 6 $ ; Ag/Dev $ r_{\text{boot}}^{2} = 0. 7 0 $ ). The geographic applicability of bioindicators was assessed by limiting sample geographical range during model calibrations. Geographic scale was limited by creating bioindicators using samples from: (a) each river, and (b) combined Mississippi/Missouri samples excluding Ohio River sites which were chemically unique. Indicator performance decreased with geographically restrictive models, therefore river basin-wide models, developed across all three rivers, is recommended. The most effective diatom-based disturbance bioindicators for this great river ecosystem could be applied using phytoplankton, periphyton, or combined assemblages to infer both trophic and agriculture/development disturbances. 相似文献
197.
Philip Jarvis Shiela Srivastav Elvira Vogelwedde Jane Stewart Terri Mitchard Gerhard F. Weinbauer 《Birth defects research. Part B, Developmental and reproductive toxicology》2010,89(3):175-187
BACKGROUND: This work evaluates pregnancy and infant loss in 1,069 vehicle‐treated cynomolgus monkeys from 78 embryo‐fetal development (EFD) studies and 14 pre‐postnatal development (PPND) studies accrued during 1981–2007. METHODS: Losses were analysed by survival function and hazard ratio using logistic regression for influence of year, study type (e.g., dose duration), and test item route of administration (ig, im, iv, sc). RESULTS: Neither study type nor route of dosing affected pregnancy outcome. Losses were higher pre‐1990 (104 losses/347 pregnancies) compared to 1990 onwards (94 losses/722 pregnancies). Losses were greatest before gestation day 50 and at parturition. Using post‐1989 data, Monte‐Carlo simulations of pregnancy outcomes were created. The power associated with the comparison of vehicle survival curves and simulated adverse survival curves was examined. This showed that EFD studies with initial vehicle group sizes of 16 and 20 have an 80% probability of having 13 and 16 ongoing pregnancies at gestational day 100, respectively. For PPND studies with initial vehicle group sizes of 16, 20, or 28, there is an 80% likelihood of having 9, 11, or 16 infants at day 7 post‐partum, respectively. A PPND study initiated with group size 20 could detect a threefold increase of test item–related pregnancy or infant loss. CONCLUSIONS: For designing and managing primate developmental toxicity studies, this type of analysis provides an objective tool to facilitate decisions either by supplementing groups with additional pregnant animals or stopping a group because an adverse effect on offspring survival has already been adequately revealed. Birth Defects Res (Part B) 89:175–187, 2010.© 2010 Wiley‐Liss, Inc. 相似文献
198.
Xiaoyu Wang Susan V. McLennan Terri J. Allen Stephen M. Twigg 《Journal of cell communication and signaling》2010,4(1):15-23
Connective tissue growth factor (CTGF), also known as CCN2, is implicated in fibrosis through both extracellular matrix (ECM)
induction and inhibition of ECM degradation. The role of CTGF in inflammation in cardiomyocytes is unknown. In some mesenchymal
cell systems, CTGF mediates effects through TGF-β or tyrosine kinase cell surface receptor, TrkA, signalling. In this study,
cellular mechanisms by which CTGF regulates pathways involved in fibrosis and inflammation were explored. Murine H9c2 cardiomyocytes
were treated with recombinant human (rh)CTGF and ECM formation gene expression: fibronectin, collagen type -I and -III and
ECM degradation genes: TIMP-1, TIMP-2 and PAI-1 were found to be induced. CTGF treatment also increased pro-inflammatory cytokines
TNF-α, IL-6, MCP-1 and IL-8. CTGF upregulated TGF-β1 mRNA and rapidly induced phosphorylation of TrkA. The CTGF-induced pro-fibrotic
and pro-inflammatory effects were blocked by anti-TGF-β neutralizing antibody and Alk 5 inhibitor (SB431542). A specific blocker
of TrkA activation, k252a, also abrogated CTGF-induced effects on fibrosis and gene expresison of MCP-1 and IL-8, but not
TNF-α or IL-6. Collectively, this data implicates CTGF in effects on pro-fibrotic genes and pro-inflammatory genes via TGF-β
pathway signalling and partly through TrkA. 相似文献
199.
Meredith H. Shaffer Yanping Huang Evann Corbo Gregory F. Wu Marielena Velez John K. Choi Ichiko Saotome Judy L. Cannon Andrea I. McClatchey Anne I. Sperling Jonathan S. Maltzman Paula M. Oliver Avinash Bhandoola Terri M. Laufer Janis K. Burkhardt 《PloS one》2010,5(8)
Background
Ezrin/radixin/moesin (ERM) proteins are highly homologous proteins that function to link cargo molecules to the actin cytoskeleton. Ezrin and moesin are both expressed in mature lymphocytes, where they play overlapping roles in cell signaling and polarity, but their role in lymphoid development has not been explored.Methodology/Principal Findings
We characterized ERM protein expression in lymphoid tissues and analyzed the requirement for ezrin expression in lymphoid development. In wildtype mice, we found that most cells in the spleen and thymus express both ezrin and moesin, but little radixin. ERM protein expression in the thymus was differentially regulated, such that ezrin expression was highest in immature thymocytes and diminished during T cell development. In contrast, moesin expression was low in early thymocytes and upregulated during T cell development. Mice bearing a germline deletion of ezrin exhibited profound defects in the size and cellularity of the spleen and thymus, abnormal thymic architecture, diminished hematopoiesis, and increased proportions of granulocytic precursors. Further analysis using fetal liver chimeras and thymic transplants showed that ezrin expression is dispensable in hematopoietic and stromal lineages, and that most of the defects in lymphoid development in ezrin−/− mice likely arise as a consequence of nutritional stress.Conclusions/Significance
We conclude that despite high expression in lymphoid precursor cells, ezrin is dispensable for lymphoid development, most likely due to redundancy with moesin. 相似文献200.
Regulation of human immunodeficiency virus type 1 envelope glycoprotein fusion by a membrane-interactive domain in the gp41 cytoplasmic tail 下载免费PDF全文
Wyss S Dimitrov AS Baribaud F Edwards TG Blumenthal R Hoxie JA 《Journal of virology》2005,79(19):12231-12241
Truncation of the human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) gp41 cytoplasmic tail (CT) can modulate the fusogenicity of the envelope glycoprotein (Env) on infected cells and virions. However, the CT domains involved and the underlying mechanism responsible for this "inside-out" regulation of Env function are unknown. HIV and SIV CTs are remarkably long and contain amphipathic alpha-helical domains (LLP1, LLP2, and LLP3) that likely interact with cellular membranes. Using a cell-cell fusion assay and a panel of HIV Envs with stop codons at various positions in the CT, we show that truncations of gp41 proximal to the most N-terminal alpha helix, LLP2, increase fusion efficiency and expose CD4-induced epitopes in the Env ectodomain. These effects were not seen with a truncation distal to this domain and before LLP1. Using a dye transfer assay to quantitate fusion kinetics, we found that these truncations produced a two- to fourfold increase in the rate of fusion. These results were observed for X4-, R5-, and dual-tropic Envs on CXCR4- and CCR5-expressing target cells and could not be explained by differences in Env surface expression. These findings suggest that distal to the membrane-spanning domain, an interaction of the gp41 LLP2 domain with the cell membrane restricts Env fusogenicity during Env processing. As with murine leukemia viruses, where cleavage of a membrane-interactive R peptide at the C terminus is required for Env to become fusogenic, this restriction of Env function may serve to protect virus-producing cells from the membrane-disruptive effects of the Env ectodomain. 相似文献