bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of a Role for an Azide-Sensitive Factor in the Thylakoid Transport of the 17-Kilodalton Subunit of the Photosynthetic Oxygen-Evolving Complex1
Authors: Leheny Ellen A. , Teter Sarah A. , Theg Steven M. 
Published: July 2002

Abstract
We have examined the transport of the precursor of the 17-kD subunit of the photosynthetic O2-evolving complex (OE17) in intact chloroplasts in the presence of inhibitors that block two protein-translocation pathways in the thylakoid membrane. This precursor uses the transmembrane pH gradient-dependent pathway into the thylakoid lumen, and its transport across the thylakoid membrane is thought to be independent of ATP and the chloroplast SecA homolog, cpSecA. We unexpectedly found that azide, widely considered to be an inhibitor of cpSecA, had a profound effect on the targeting of the photosynthetic OE17 to the thylakoid lumen. By itself, azide caused a significant fraction of mature OE17 to accumulate in the stroma of intact chloroplasts. When added in conjunction with the protonophore nigericin, azide caused the maturation of a fraction of the stromal intermediate form of OE17, and this mature protein was found only in the stroma. Our data suggest that OE17 may use the sec-dependent pathway, especially when the transmembrane pH gradient-dependent pathway is inhibited. Under certain conditions, OE17 may be inserted across the thylakoid membrane far enough to allow removal of the transit peptide, but then may slip back out of the translocation machinery into the stromal compartment.
Direct Evidence for Rapid Degradation ofBacillus thuringiensis Toxin mRNA as a Cause of Poor Expression in Plants1
Authors: De Rocher E. Jay , Vargo-Gogola Tracy C. , Diehn Scott H. , Green Pamela J. 
Published: July 2002

Abstract
It is well established that the expression of Bacillus thuringiensis (B.t.) toxin genes in higher plants is severely limited at the mRNA level, but the cause remains controversial. Elucidating whether mRNA accumulation is limited transcriptionally or posttranscriptionally could contribute to effective gene design as well as provide insights about endogenous plant gene-expression mechanisms. To resolve this controversy, we compared the expression of an A/U-rich wild-typecryIA(c) gene and a G/C-rich synthetic cryIA(c) B.t.-toxin gene under the control of identical 5′ and 3′ flanking sequences. Transcriptional activities of the genes were equal as determined by nuclear run-on transcription assays. In contrast, mRNA half-life measurements demonstrated directly that the wild-type transcript was markedly less stable than that encoded by the synthetic gene. Sequences that limit mRNA accumulation were located at more than one site within the coding region, and some appeared to be recognized in Arabidopsis but not in tobacco (Nicotiana tabacum). These results support previous observations that some A/U-rich sequences can contribute to mRNA instability in plants. Our studies further indicate that some of these sequences may be differentially recognized in tobacco cells and Arabidopsis.
Proteasome Inhibitors Prevent Tracheary Element Differentiation in Zinnia Mesophyll Cell Cultures
Authors: Woffenden Bonnie J. , Freeman Thomas B. , Beers Eric P. 
Published: July 2002

Abstract
To determine whether proteasome activity is required for tracheary element (TE) differentiation, the proteasome inhibitors clasto-lactacystin β-lactone and carbobenzoxy-leucinyl-leucinyl-leucinal (LLL) were used in a zinnia (Zinnia elegans) mesophyll cell culture system. The addition of proteasome inhibitors at the time of culture initiation prevented differentiation otherwise detectable at 96 h. Inhibition of the proteasome at 48 h, after cellular commitment to differentiation, did not alter the final percentage of TEs compared with controls. However, proteasome inhibition at 48 h delayed the differentiation process by approximately 24 h, as indicated by examination of both morphological markers and the expression of putative autolytic proteases. These results indicate that proteasome function is required both for induction of TE differentiation and for progression of the TE program in committed cells. Treatment at 48 h with LLL but not clasto-lactacystin β-lactone resulted in partial uncoupling of autolysis from differentiation. Results from gel analysis of protease activity suggested that the observed incomplete autolysis was due to the ability of LLL to inhibit TE cysteine proteases.
A Gene Encoding Proline Dehydrogenase Is Not Only Induced by Proline and Hypoosmolarity, but Is Also Developmentally Regulated in the Reproductive Organs of Arabidopsis
Authors: Nakashima Kazuo , Satoh Rie , Kiyosue Tomohiro , Yamaguchi-Shinozaki Kazuko , Shinozaki Kazuo 
Published: July 2002

AbstractThe cDNA clone ERD5 (earlyresponsive to dehydration), isolated from 1-h-dehydrated Arabidopsis, encodes a precursor of proline (Pro) dehydrogenase (ProDH), which is a mitochondrial enzyme involved in the first step of the conversion of Pro to glutamic acid. The transcript of the erd5 (ProDH) gene was undetectable when plants were dehydrated, but large amounts of transcript accumulated when plants were subsequently rehydrated. Accumulation of the transcript was also observed in plants that had been incubated under hypoosmotic conditions in media that contained l- ord-Pro. We isolated a 1.4-kb DNA fragment of the putative promoter region of the ProDH gene. Theβ-glucuronidase (GUS) reporter gene driven by the 1.4-kb ProDH promoter was induced not only by rehydration but also by hypoosmolarity and l- andd-Pro at significant levels in transgenic Arabidopsis plants. The promoter of the ProDH gene directs strong GUS activity in reproductive organs such as pollen and pistils and in the seeds of the transgenic plants. GUS activity was detected in vegetative tissues such as veins of leaves and root tips when the transgenic plants were exposed to hypoosmolarity and Pro solutions. GUS activity increased during germination of the transgenic plants under hypoosmolarity. The relationship between Pro metabolism and the physiological aspects of stress response and development are discussed.
High-Molecular-Weight FK506-Binding Proteins Are Components of Heat-Shock Protein 90 Heterocomplexes in Wheat Germ Lysate
Authors: Reddy Ramachandra K. , Kurek Isaac , Silverstein Adam M. , Chinkers Michael , Breiman Adina , Krishna Priti 
Published: July 2002

Abstract
In animal cell lysates the multiprotein heat-shock protein 90 (hsp90)-based chaperone complexes consist of hsp70, hsp40, and p60. These complexes act to convert steroid hormone receptors to their steroid-binding state by assembling them into heterocomplexes with hsp90, p23, and one of several immunophilins. Wheat germ lysate also contains a hsp90-based chaperone system that can assemble the glucocorticoid receptor into a functional heterocomplex with hsp90. However, only two components of the heterocomplex-assembly system, hsp90 and hsp70, have thus far been identified. Recently, purified mammalian p23 preadsorbed with JJ3 antibody-protein A-Sepharose pellets was used to isolate a mammalian p23-wheat hsp90 heterocomplex from wheat germ lysate (J.K. Owens-Grillo, L.F. Stancato, K. Hoffmann, W.B. Pratt, and P. Krishna [1996] Biochemistry 35: 15249–15255). This heterocomplex was found to contain an immunophilin(s) of the FK506-binding class, as judged by binding of the radiolabeled immunosuppressant drug [3H]FK506 to the immune pellets in a specific manner. In the present study we identified the immunophilin components of this heterocomplex as FKBP73 and FKBP77, the two recently described high-molecular-weight FKBPs of wheat. In addition, we present evidence that the two FKBPs bind hsp90 via tetratricopeptide repeat domains. Our results demonstrate that binding of immunophilins to hsp90 via tetratricopeptide repeat domains is a conserved protein interaction in plants. Conservation of this protein-to-protein interaction in both plant and animal cells suggests that it is important for the biological action of the high-molecular-weight immunophilins.
Regulation of Soybean Nodulation Independent of Ethylene Signaling1
Authors: Schmidt J. Scott , Harper James E. , Hoffman Thomas K. , Bent Andrew F. 
Published: July 2002

Abstract
Leguminous plants regulate the number of Bradyrhizobium- or Rhizobium-infected sites that develop into nitrogen-fixing root nodules. Ethylene has been implicated in the regulation of nodule formation in some species, but this role has remained in question for soybean (Glycine max). The present study used soybean mutants with decreased responsiveness to ethylene, soybean mutants with defective regulation of nodule number, and Ag+ inhibition of ethylene perception to examine the role of ethylene in the regulation of nodule number. Nodule numbers on ethylene-insensitive mutants and plants treated with Ag+ were similar to those on wild-type plants and untreated plants, respectively. Hypernodulating mutants displayed wild-type ethylene sensitivity. Suppression of nodule numbers by high nitrate was also similar between ethylene-insensitive plants, wild-type plants, and plants treated with Ag+. Ethylene insensitivity of the roots of etr1-1 mutants was confirmed using assays for sensitivity to 1-aminocyclopropane-1-carboxylic acid and for ethylene-stimulated root-hair formation. Additional phenotypes ofetr1-1 roots were also characterized. Ethylene-dependent pathways regulate the number of nodules that form on species such as pea and Medicago truncatula, but our data indicate that ethylene is less significant in regulating the number of nodules that form on soybean.
Prediction of Protein Cleavage Sites by the Barley Cysteine Endoproteases EP-A and EP-B Based on the Kinetics of Synthetic Peptide Hydrolysis
Authors: Davy Anne , Sørensen Mikael Blom , Svendsen Ib , Cameron-Mills Verena , Simpson David J. 
Published: July 2002

Abstract
Hordeins, the natural substrates of barley (Hordeum vulgare) cysteine endoproteases (EPs), were isolated as protein bodies and degraded by purified EP-B from green barley malt. Cleavage specificity was determined by synthesizing internally quenched, fluorogenic tetrapeptide substrates of the general formula 2-aminobenzoyl-P2-P1-P1′-P2′ 1-tyrosine(NO2)-aspartate. The barley EPs preferred neutral amino acids with large aliphatic and nonpolar (leucine, valine, isoleucine, and methionine) or aromatic (phenylalanine, tyrosine, and tryptophan) side chains at P2, and showed less specificity at P1, although asparagine, aspartate, valine, and isoleucine were particularly unfavorable. Peptides with proline at P1 or P1′ were extremely poor substrates. Cleavage sites with EP-A and EP-B preferred substrate sequences are found in hordeins, their natural substrates. The substrate specificity of EP-B with synthetic peptides was used successfully to predict the cleavage sites in the C-terminal extension of barley β-amylase. When all of the primary cleavage sites in C hordein, which occur mainly in the N- and C-terminal domains, were removed by site-directed mutagenesis, the resulting protein was degraded 112 times more slowly than wild-type C hordein. We suggest that removal of the C hordein terminal domains is necessary for unfolding of the β-reverse turn helix of the central repeat domain, which then becomes more susceptible to proteolytic attack by EP-B.
Spatio-Temporal Accumulation and Activity of Calcium-Dependent Protein Kinases during Embryogenesis, Seed Development, and Germination in Sandalwood
Authors: Anil Veena S. , Harmon Alice C. , Rao K. Sankara 
Published: July 2002

Abstract
Western-blot analysis and protein kinase assays identified two Ca2+-dependent protein kinases (CDPKs) of 55 to 60 kD in soluble protein extracts of embryogenic cultures of sandalwood (Santalum album L.). However, these sandalwood CDPKs (swCDPKs) were absent in plantlets regenerated from somatic embryos. swCDPKs exhibited differential expression (monitored at the level of the protein) and activity in different developmental stages. Zygotic embryos, seedlings, and endosperm showed high accumulation of swCDPK, but the enzyme was not detected in the soluble proteins of shoots and flowers. swCDPK exhibited a temporal pattern of expression in endosperm, showing high accumulation and activity in mature fruit and germinating stages; the enzyme was localized strongly in the storage bodies of the endosperm cells. The study also reports for the first time to our knowledge a post-translational inhibition/inactivation of swCDPK in zygotic embryos during seed dormancy and early stages of germination. The temporal expression of swCDPK during somatic/zygotic embryogenesis, seed maturation, and germination suggests involvement of the enzyme in these developmental processes.
Ascorbate Biosynthesis in Mitochondria Is Linked to the Electron Transport Chain between Complexes III and IV
Authors: Bartoli Carlos G. , Pastori Gabriela M. , Foyer Christine H. 
Published: July 2002

Abstract
Ascorbic acid is synthesized from galactono-γ-lactone (GL) in plant tissues. An improved extraction procedure involving ammonium sulfate precipitation of membrane proteins from crude leaf homogenates yielded a simple, quick method for determining tissue activities of galactono-γ-lactone dehydrogenase (GLDH). Total foliar ascorbate and GLDH activity decreased with leaf age. Subcellular fractionation experiments using marker enzymes demonstrated that 80% of the total GLDH activity was located on the inner mitochondrial membrane, and 20% in the microsomal fraction. Specific antibody raised against potato (Solanum tuberosum L.) tuber GLDH recognized a 56-kD polypeptide in extracts from the mitochondrial membranes but failed to detect the equivalent polypeptide in microsomes. We demonstrate that isolated intact mitochondria synthesize ascorbate in the presence of GL. GL stimulated mitochondrial electron transport rates. The respiration inhibitor antimycin A stimulated ascorbate biosynthesis, while cyanide inhibited both respiration and ascorbate production. GL-dependent oxygen uptake was observed in isolated intact mitochondria. This evidence suggests that GLDH delivers electrons to the mitochondrial electron transport chain between complexes III and IV.
Biochemical Characterization of Wild-Type and Mutant Isoamylases of Chlamydomonas reinhardtii Supports a Function of the Multimeric Enzyme Organization in Amylopectin Maturation
Authors: Dauvillée David , Colleoni Christophe , Mouille Gregory , Morell Matthew K. , d'Hulst Christophe , Wattebled Fabrice , Liénard Luc , Delvallé David , Ral Jean-Philippe , Myers Alan M. , Ball Steven G. 
Published: July 2002

Abstract
Chlamydomonas reinhardtii mutants of theSTA8 gene produce reduced amounts of high amylose starch and phytoglycogen. In contrast to the previously described phytoglycogen-producing mutants of C. reinhardtii that contain no residual isoamylase activity, the sta8mutants still contained 35% of the normal amount of enzyme activity. We have purified this residual isoamylase and compared it with the wild-type C. reinhardtii enzyme. We have found that the high-mass multimeric enzyme has reduced its average mass at least by one-half. This coincides with the disappearance of two out of the three activity bands that can be seen on zymogram gels. Wild-type and mutant enzymes are shown to be located within the plastid. In addition, they both act by cleaving off the outer branches of polysaccharides with no consistent difference in enzyme specificity. Because the mutant enzyme was demonstrated to digest phytoglycogen to completion in vitro, we propose that its inability to do so in vivo supports a function of the enzyme complex architecture in the processing of pre-amylopectin chains.
The Sucrose Analog Palatinose Leads to a Stimulation of Sucrose Degradation and Starch Synthesis When Supplied to Discs of Growing Potato Tubers
Authors: Fernie Alisdair R. , Roessner Ute , Geigenberger Peter 
Published: July 2002

Abstract
In the present paper we investigated the effect of the sucrose (Suc) analog palatinose on potato (Solanum tuberosum) tuber metabolism. In freshly cut discs of growing potato tubers, addition of 5 mm palatinose altered the metabolism of exogenously supplied [U-14C]Suc. There was slight inhibition of the rate of 14C-Suc uptake, a 1.5-fold increase in the rate at which 14C-Suc was subsequently metabolized, and a shift in the allocation of the metabolized label in favor of starch synthesis. The sum result of these changes was a 2-fold increase in the absolute rate of starch synthesis. The increased rate of starch synthesis was accompanied by a 3-fold increase in inorganic pyrophosphate, a 2-fold increase in UDP, decreased UTP/UDP, ATP/ADP, and ATP/AMP ratios, and decreased adenylate energy charge, whereas glycolytic and Krebs cycle intermediates were unchanged. In addition, feeding palatinose to potato discs also stimulated the metabolism of exogenous 14C-glucose in favor of starch synthesis. In vitro studies revealed that palatinose is not metabolized by Suc synthases or invertases within potato tuber extracts. Enzyme kinetics revealed different effects of palatinose on Suc synthase and invertase activities, implicating palatinose as an allosteric effector leading to an inhibition of Suc synthase and (surprisingly) to an activation of invertase in vitro. However, measurement of tissue palatinose levels revealed that these were too low to have significant effects on Suc degrading activities in vivo. These results suggest that supplying palatinose to potato tubers represents a novel way to increase starch synthesis.
The Delayed Terminal Flower Phenotype Is Caused by a Conditional Mutation in the CENTRORADIALIS Gene of Snapdragon
Authors: Cremer Frédéric , Lönnig Wolf-Ekkehard , Saedler Heinz , Huijser Peter 
Published: July 2002

Abstract
The snapdragon (Antirrhinum majus)centroradialis mutant (cen) is characterized by the development of a terminal flower, thereby replacing the normally open inflorescence by a closed inflorescence. In contrast to its Arabidopsis counterpart, terminal flower1, the cen-null mutant displays an almost constant number of lateral flowers below the terminal flower. Some partial revertants of an X-radiation-induced cen mutant showed a delayed formation of the terminal flower, resulting in a variable number of lateral flowers. The number of lateral flowers formed was shown to be environmentally controlled, with the fewer flowers formed under the stronger flower-inducing conditions. Plants displaying this “Delayed terminal flower” phenotype were found to be heterozygous for a mutant allele carrying a transposon in the coding region and an allele from which the transposon excised, leaving behind a 3-bp duplication as footprint. As a consequence, an iso-leucine is inserted between Asp148 and Gly149 in the CENTRORADIALIS protein. It is proposed that this mutation results in a low level of functional CEN activity, generating a phenotype that is more similar to the Arabidopsis Terminal flower phenotype.
Proton and metal ion‐dependent assembly of a model diiron protein
Authors: Pasternak Annette , Kaplan Justin , Lear James D. , Degrado William F. 
Published: July 2002

AbstractDF1 is a small, idealized model for carboxylate‐bridged diiron proteins. This protein was designed to form a dimeric four‐helix bundle with a dimetal ion‐binding site near the center of the structure, and its crystal structure has confirmed that it adopts the intended conformation. However, the protein showed limited solubility in aqueous buffer, and access to its active site was blocked by two hydrophobic side chains. The sequence of DF1 has now been modified to provide a very soluble protein (DF2) that binds metal ions in a rapid and reversible manner. Furthermore, the DF2 protein shows significant ferroxidase activity, suggesting that its dimetal center is accessible to oxygen. The affinity of DF2 for various first‐row divalent cations deviates from the Irving–Willliams series, suggesting that its structure imparts significant geometric preferences on the metal ion‐binding site. Furthermore, in the absence of metal ions, the protein folds into a dimer with concomitant binding of two protons. The uptake of two protons is expected if the structure of the apo‐protein is similar to that of the crystal structure of dizinc DF1. Thus, this result suggests that the active site of DF2 is retained in the absence of metal ions.
Mean‐Variance Hedging and Numéraire
Authors: Gourieroux Christian , Laurent Jean Paul , Pham Huyên 
Published: July 2002

We consider the mean‐variance hedging problem when the risky assets price process is a continuous semimartingale. The usual approach deals with self‐financed portfolios with respect to the primitive assets family. By adding a numéraire as an asset to trade in, we show how self‐financed portfolios may be expressed with respect to this extended assets family, without changing the set of attainable contingent claims.We introduce the hedging numéraire and relate it to the variance‐optimal martingale measure. Using this numéraire both as a deflator and to extend the primitive assets family, we are able to transform the original mean‐variance hedging problem into an equivalent and simpler one; this transformed quadratic optimization problem is solved by the Galtchouk–Kunita–Watanabe projection theorem under a martingale measure for the hedging numéraire extended assets family. This gives in turn an explicit description of the optimal hedging strategy for the original mean‐variance hedging problem.
Design of Robust Vibration Controller for a Smart Panel Using Finite
Element Model
Authors: Chang W. , Gopinathan Senthil V. , Varadan V. V. , Varadan V. K. 
Published: July 2002

Abstract
This paper presents a model reduction method and uncertainty modeling for the design of a low-order H∞ robust controller for suppression of smart panel vibration. A smart panel with collocated piezoceramic actuators and sensors is modeled using solid, transition, and shell finite elements, and then the size of the model is reduced in the state space domain. A robust controller is designed not only to minimize the panel vibration excited by applied uniform acoustic pressure, but also to be reliable in real world applications. This paper introduces the idea of Modal Hankel Singular values (MHSV) to reduce the finite element model to a low-order state space model with minimum model reduction error. MHSV measures balanced controllability and observability of each resonance mode to deselect insignificant resonance modes. State space modeling of realistic control conditions are formulated in terms of uncertainty variables. These uncertainty variables include uncertainty in actuators and sensors performances, uncertainty in the knowledge of resonance frequencies of the structure, damping ratio, static stiffness, unmodeled high resonance vibration modes, etc. The simplified model and the uncertainty model are combined as an integrated state space model, and then implemented in the H∞ control theory for controller parameterization. The low-order robust controller is easy to implement in an analog circuit to provide a low cost solution in a variety of applications where cost may be a limiting factor.