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921.
922.
923.
Mahima Kaushik Manoj Prasad Shikha Kaushik Anju Singh Shrikant Kukreti 《Biopolymers》2010,93(2):150-160
Widely dispersed in genomic DNA, the tandem C‐rich repetitive stretches may fold below physiological pH, into i‐motif structures, stabilized by C·C+ pairing. Herein, structural status of a 9‐mer stretch d(CCCTAACCC), [the truncated double repeat of human telomeric sequence], and its extended version, comprising of additional ? TAA segment at the 3′‐end, representing the complete double repeat d(CCCTAACCCTAA), has been investigated. The pH dependent monophasic UV‐melting, Gel and CD data suggested that while the truncated version adopts a bimolecular i‐motif structure, its complete double repeat (12‐mer) sequence exists in two (bimolecular and tetramolecular) forms. A model is proposed for the tetramolecular i‐motif with conventional C · C+ base pairs, additionally stabilized by asymmetric A · A base pairs at the ?3′ TAA flanking ends and Watson–Crick A · T hydrogen bonding between intervening bases on antiparallel strands. Expanding the known topologies of DNA i‐motifs, such atypical geometries of i‐motifs may have implications in their recognition by proteins. © 2009 Wiley Periodicals, Inc. Biopolymers 93: 150–160, 2010. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com 相似文献
924.
Robert J. Ossiboff Yi Zhou Patrick J. Lightfoot B. V. Venkataram Prasad John S. L. Parker 《Journal of virology》2010,84(11):5550-5564
Nonenveloped viral capsids are metastable structures that undergo conformational changes during virus entry that lead to interactions of the capsid or capsid fragments with the cell membrane. For members of the Caliciviridae, neither the nature of these structural changes in the capsid nor the factor(s) responsible for inducing these changes is known. Feline junctional adhesion molecule A (fJAM-A) mediates the attachment and infectious viral entry of feline calicivirus (FCV). Here, we show that the infectivity of some FCV isolates is neutralized following incubation with the soluble receptor at 37°C. We used this property to select mutants resistant to preincubation with the soluble receptor. We isolated and sequenced 24 soluble receptor-resistant (srr) mutants and characterized the growth properties and receptor-binding activities of eight mutants. The location of the mutations within the capsid structure of FCV was mapped using a new 3.6-Å structure of native FCV. The srr mutations mapped to the surface of the P2 domain were buried at the protruding domain dimer interface or were present in inaccessible regions of the capsid protein. Coupled with data showing that both the parental FCV and the srr mutants underwent increases in hydrophobicity upon incubation with the soluble receptor at 37°C, these findings indicate that FCV likely undergoes conformational change upon interaction with its receptor. Changes in FCV capsid conformation following its interaction with fJAM-A may be important for subsequent interactions of the capsid with cellular membranes, membrane penetration, and genome delivery.The interactions between viruses and receptors on the surface of host cells strongly influence viral pathogenesis and regulate morbidity and mortality in the host. Virus-receptor interactions determine the types of cells that can be infected, the pathway of entry into the cell, and the efficiency of productive infection. Interactions between nonenveloped virus capsids and their receptor(s) trigger one or more steps required for infectious entry. These steps can include interaction with other receptors, exposure to low pH or endosomal proteases, or other factors. Ultimately, one or more of these interactions induce changes in capsid conformation that result in the exposure of hydrophobic regions or release of a lipid-seeking factor that can interact with and disrupt the limiting cellular membrane to allow the capsid and/or the genome to be delivered to the interior of the cell (reviewed in reference 60).The Caliciviridae are small nonenveloped viruses containing a positive-sense RNA genome (∼7 to 8 kb). Several important disease-causing members of the Caliciviridae, including human noroviruses and rabbit hemorrhagic disease virus, cannot be propagated in tissue culture systems (19, 56). This has slowed progress on studies of the mechanisms of cellular entry of these viruses. In contrast, feline caliciviruses (FCVs) propagate readily in tissue culture, and two cell surface receptor molecules, feline junctional adhesion molecule A (fJAM-A) and α2,6 sialic acid, are known (29, 55).The FCV receptor, fJAM-A, is a type I transmembrane glycoprotein (molecular size of 36 to 41 kDa) member of the immunoglobulin superfamily (IgSF); it consists of an amino-terminal signal peptide, an extracellular domain (composed of two Ig-like domains—a membrane-distal D1 and a membrane-proximal D2), a transmembrane domain, and a short cytoplasmic domain that contains a type II PDZ domain-binding motif (11, 30). We have previously shown that the D1 domain of the fJAM-A ectodomain is necessary and sufficient for FCV binding and that preincubation of FCV with soluble fJAM-A (sfJAM-A) results in virus neutralization (35). Additional roles that fJAM-A might play in FCV entry, however, have not been investigated.Caliciviruses are composed of 180 copies of a single capsid protein. Atomic resolution structures of recombinant virus-like particles of Norwalk virus (genus Norovirus) and native San Miguel sea lion virus (SMSV) virions (genus Vesivirus) indicate that the virion consists of 90 dimers of the capsid protein arranged in T=3 icosahedral symmetry (5, 41). Each capsid monomer contains three structural domains—an N-terminal arm (NTA), the shell (S), and a protruding domain (P) that is further subdivided into P1 and P2 subdomains. The distal subdomain, P2, is structurally conserved between Norwalk virus and SMSV, but there is little sequence conservation. In the primary sequence of the FCV capsid, there are two hypervariable regions that contain neutralizing epitopes (18, 34, 58). The corresponding hypervariable regions (HVRs) of the SMSV capsid structure map to surface-exposed loops. Surface residues at the dimeric interface between two capsid monomers are conserved within individual calicivirus genera, and it has been suggested that this interface is involved in receptor binding (5). A cryo-electron microscopy (cryo-EM) reconstruction of the FCV vaccine strain F9 complexed with the ectodomain of fJAM-A (modeled on the crystal structures of SMSV and human JAM-A, respectively) shows that fJAM-A engages the top of the P2 domain and that binding causes a rotation in the P dimer (1). However, the relatively low resolution and the lack of atomic resolution structures of FCV and fJAM-A prevented precise identification of residues on the viral capsid that contact fJAM-A.A classical approach for identifying virus residues that directly bind or modulate the binding of a receptor is to select for mutant viruses resistant to neutralization with soluble receptors (6, 23, 46). Soluble receptor-resistant (srr) mutants of poliovirus revealed that both surface-exposed and internal residues regulate receptor attachment and conformational changes in the capsid (6, 42). Here, we report 24 srr mutants and the location of their capsid mutations on a 3.6-Å structure of FCV. In addition, we describe the growth kinetics and receptor-binding properties of a subpanel of eight srr mutants and examine changes in capsid hydrophobicity concurrent with the interaction of FCV capsids with sfJAM-A. 相似文献
925.
P. Rama Chandra Prasad K. S. Rajan C. B. S. Dutt P. S. Roy 《Biodiversity and Conservation》2010,19(11):3073-3087
Phytodiversity is affected both by natural and anthropogenic factors and in Island ecosystems these impacts can devastate
or reduce diversity, if the native vegetation is lost. In addition to rich species richness and diversity, Island systems
are the sites of high endemism and any threat to these ecosystems will consequently lead to loss and extinction of species.
To understand the dynamics including feedbacks of these changes in phytodiversity of North Andaman Islands, a conceptual framework
is proposed which focuses on understanding the land-use and land-cover changes and its impact with phytodiversity. In considering
land-use and land-cover changes this work highlights the direct and indirect drivers of changes—socio-economic, biophysical
and climatic factors. Migration of population, their socio economic needs and government policies were identified as major
driving forces threatening the phytodiversity of these Islands. Apart from human beings, natural disasters like tsunami and
introduced herbivorous animals like elephants also contributed to forest destruction in these Islands. The integrated analysis
based on such framework will provide insights for holistic resource management including ecological conservation. 相似文献
926.
Henrik Johansson Olov Svartström Prasad Phadnis Lars Engman Marjam Karlsson Ott 《Bioorganic & medicinal chemistry》2010,18(5):1783-1788
The organoselenium antioxidant 1 was previously found to act as a catalytic antioxidant in a two-phase lipid peroxidation system. In aqueous environment, selenide 1 quenched ABTS-radicals more efficiently than Trolox and ascorbic acid. The selenide dose-dependently scavenged reactive oxygen and nitrogen species more efficiently than Trolox for neutrophils and PMA-stimulated macrophages, with 50% inhibitory concentrations in the low micromolar range. In addition no sign of toxicity or effect on cell viability was seen when culturing five human cell lines in concentrations up to 200 μM of selenide 1 for up to seven days. We therefore feel that the compound would be a good candidate for future drug development for prevention or treatment of disorders caused by or involving free radical-mediated or oxidative tissue damage. 相似文献
927.
928.
Bhaskarjyoti Prasad Shardul S. Salunkhe Sriram Padmanabhan 《Protein expression and purification》2010,69(2):191-197
Staphylokinase (SAK) is reported to have a serine protease domain with no proteolytic activity unlike other plasminogen activators like tissue plasminogen activator (t-PA) and urokinase. A unique protease property of Staphylokinase was observed when SAK was expressed as a fusion protein in inducible Escherichia coli expression vectors. This finding was further investigated by cloning and expressing different SAK fusions, both native and N-terminal deletions, with fusion tags like glutathione S-transferase (GST) and signal sequence of SAK in bacterial system. While all the N-terminal SAK fusions were found to self-cleave in crude and purified preparations, the C-terminal SAK fusion was stable. The cleavage property of Staphylokinase fusion proteins, inhibited by reduced glutathione and PMSF, was independent of its thrombolytic activity and also independent on the type of host employed for its expression. The serine protease domain of the SAK gene possibly lies between 20th to 77th amino acid and serine 41 of this region appears critical for such a cleavage property. 相似文献
929.
V. Channamallikarjuna H. Sonah M. Prasad G. J. N. Rao S. Chand H. C. Upreti N. K. Singh T. R. Sharma 《Molecular breeding : new strategies in plant improvement》2010,25(1):155-166
Sheath blight caused by Rhizoctonia solani Kühn is one of the important diseases of rice, resulting in heavy yield loss in rice every year. No rice line resistant to
sheath blight has been identified till date. However, in some rice lines a high degree of resistance to R. solani has been observed. An indica rice line, Tetep, is a well documented source of durable and broad spectrum resistance to rice
blast as well as quantitative resistance to sheath blight. The present study identified genetic loci for quantitative resistance
to sheath blight in rice line Tetep. A mapping population consisting of 127 recombinant inbred lines derived from a cross
between rice cultivars HP2216 (susceptible) and Tetep (resistant to sheath blight) was evaluated for sheath blight resistance
and other agronomic traits for 4 years across three locations. Based on sheath blight phenotypes and genetic map with 126
evenly distributed molecular markers, a quantitative trait loci (QTLs) contributing to sheath blight resistance was identified
on long arm of chromosome 11. Two QTL mapping approaches i.e., single marker analysis and composite interval mapping in multi
environments were used to identify QTLs for sheath blight resistance and agronomical traits. The QTL qSBR11-1 for sheath blight
resistance was identified between the marker interval RM1233 (26.45 Mb) to sbq33 (28.35 Mb) on chromosome 11. This region
was further narrowed down to marker interval K39516 to sbq33 (~0.85 Mb) and a total of 154 genes were predicted including
11 tandem repeats of chitinase genes which may be responsible for sheath blight resistance in rice line Tetep. A set of 96
varieties and a F2 population were used for validation of markers linked to the QTL region. The results indicate that there is very high genetic
variation among varieties at this locus, which can serve as a starting point for allele mining of sheath blight resistance. 相似文献
930.
Meenakshi Gaur Carissa Ritner Rich Sievers Anissa Pedersen Megha Prasad Harold S. Bernstein Yerem Yeghiazarians 《Cytotherapy》2010,12(6):807-817
Background aimsHeart failure therapy with human embryonic stem cell (hESC)-derived cardiomyocytes (hCM) has been limited by the low rate of spontaneous hCM differentiation. As others have shown that p38 mitogen-activated protein kinase (p38MAPK) directs neurogenesis from mouse embryonic stem cells, we investigated whether the p38MAPK inhibitor, SB203580, might influence hCM differentiation.MethodsWe treated differentiating hESC with SB203580 at specific time-points, and used flow cytometry, immunocytochemistry, quantitative real-time (RT)–polymerase chain reaction (PCR), teratoma formation and transmission electron microscopy to evaluate cardiomyocyte formation.ResultsWe observed that the addition of inhibitor resulted in 2.1-fold enrichment of spontaneously beating human embryoid bodies (hEB) at 21 days of differentiation, and that 25% of treated cells expressed cardiac-specific α-myosin heavy chain. This effect was dependent on the stage of differentiation at which the inhibitor was introduced. Immunostaining and teratoma formation assays demonstrated that the inhibitor did not affect hESC pluripotency; however, treated hESC gave rise to hCM exhibiting increased expression of sarcomeric proteins, including cardiac troponin T, myosin light chain and α-myosin heavy chain. This was consistent with significantly increased numbers of myofibrillar bundles and the appearance of nascent Z-bodies at earlier time-points in treated hCM. Treated hEB also demonstrated a normal karyotype by array comparative genomic hybridization and viability in vivo following injection into mouse myocardium.ConclusionsThese studies demonstrate that p38MAPK inhibition accelerates directed hCM differentiation from hESC, and that this effect is developmental stage-specific. The use of this inhibitor should improve our ability to generate hESC-derived hCM for cell-based therapy. 相似文献