The fate of the initiator tRNAs is sensitive to the critical balance between interacting proteins

Formylation of the initiator tRNA is essential for normal growth of Escherichia coil, The initiator tRNA containing the U35A36 mutation (CUA anticodon) initiates from UAG codon, However, an additional mutation at position 72 (72A --> G) renders the tRNA (G72/U35A36) inactive in initiation because it is defective in formylation, In this study, we isolated U1G72/U35A36 tRNA containing a wobble base pair at 1-72 positions as an intragenic suppressor of the G72 mutation. The U1G72/U35A36 tRNA is formylated and participates in initiation. More importantly, we show that the mismatch at 1-72 positions of the initiator tRNA, which was thus far thought to be the hallmark of the resistance of this tRNA against peptidyl-tRNA hydrolase (PTH), is not sufficient, The amino acid attached to the initiator tRNA is also important in conferring protection against PTH. Further, we show that the relative levels of PTH and IF2 influence the path adopted by the initiator tRNAs in protein synthesis. These findings provide an important clue to understand the dual function of the single tRNA(Met) in initiation and elongation, in the mitochondria of various organisms.

Conformational changes of the chaperone SecB upon binding to a model substrate - bovine pancreatic trypsin inhibitor (BPTI)

SecB is a homotetrameric cytosolic chaperone that forms part of the protein translocation machinery in E. coli. Due to SecB, nascent polypeptides are maintained in an unfolded translocation-competent state devoid of tertiary structure and thus are guided to the translocon. In vitro SecB rapidly binds to a variety of ligands in a non-native state. We have previously investigated the bound state conformation of the model substrate bovine pancreatic trypsin inhibitor (BPTI) as well as the conformation of SecB itself by using proximity relationships based on site-directed spin labeling and pyrene fluorescence methods. It was shown that SecB undergoes a conformational change during the process of substrate binding. Here, we generated SecB mutants containing but a single cysteine per subunit or an exposed highly reactive new cysteine after removal of the nearby intrinsic cysteines. Quantitative spin labeling was achieved with the methanethiosulfonate spin label (MTS) at positions C97 or E90C, respectively. Highfield (W-band) electron paramagnetic resonance (EPR) measurements revealed that with BPTI present the spin labels are exposed to a more polar/hydrophilic environment. Nanoscale distance measurements with double electron-electron resonance (DEER) were in excellent agreement with distances obtained by molecular modeling. Binding of BPTI also led to a slight change in distances between labels at C97 but not at E90C. While the shorter distance in the tetramer increased, the larger diagonal distance decreased. These findings can be explained by a widening of the tetrameric structure upon substrate binding much like the opening of two pairs of scissors.

Extension of the universal erosive burning law to partly symmetric propellant grain geometries

This paper aims at extending the universal erosive burning law developed by two of the present authors from axi-symmetric internally burning grains to partly symmetric burning grains. This extension revolves around three dimensional flow calculations inside highly loaded grain geometry and benefiting from an observation that the flow gradients normal to the surface in such geometries have a smooth behavior along the perimeter of the grain. These are used to help identify the diameter that gives the same perimeter the characteristic dimension rather than a mean hydraulic diameter chosen earlier. The predictions of highly loaded grains from the newly chosen dimension in the erosive burning law show better comparison with measured pressure-time curves while those with mean hydraulic diameter definitely over-predict the pressures. (c) 2013 IAA. Published by Elsevier Ltd. All rights reserved.

Inactivation of the Bacterial RNA Polymerase Due to Acquisition of Secondary Structure by the omega Subunit

The widely conserved omega subunit encoded by rpoZ is the smallest subunit of Escherichia coli RNA polymerase (RNAP) but is dispensable for bacterial growth. Function of omega is known to be substituted by GroEL in omega-null strain, which thus does not exhibit a discernable phenotype. In this work, we report isolation of omega variants whose expression in vivo leads to a dominant lethal phenotype. Studies show that in contrast to omega, which is largely unstructured, omega mutants display substantial acquisition of secondary structure. By detailed study with one of the mutants, omega(6) bearing N60D substitution, the mechanism of lethality has been deciphered. Biochemical analysis reveals that omega(6) binds to beta ` subunit in vitro with greater affinity than that of omega. The reconstituted RNAP holoenzyme in the presence of omega(6) in vitro is defective in transcription initiation. Formation of a faulty RNAP in the presence of mutant omega results in death of the cell. Furthermore, lethality of omega(6) is relieved in cells expressing the rpoC2112 allele encoding beta ` (2112), a variant beta ` bearing Y457S substitution, immediately adjacent to the beta ` catalytic center. Our results suggest that the enhanced omega(6)-beta ` interaction may perturb the plasticity of the RNAP active center, implicating a role for omega and its flexible state.

USEFUL EXTENSIONS OF THE HENRY REACTION: EXPEDITIOUS ROUTES TO NITROALKANES AND NITROALKENES IN AQUEOUS MEDIA

The products of the Henry nitroaldol reaction from nitromethane and several aldehydes were reduced to the corresponding nitroalkanes with (n-Bu)(3)SnH in water under microwave irradiation (80 degrees C/10 min), or dehydrated to the corresponding nitroalkenes with K2CO3 in water (generally 0-5 degrees C/20 min). Both ``one-pot'' reactions occur in excellent yields across a range of aliphatic and aromatic (including heteroaromatic) substrates. It seems likely that the deoxygenation of the nitroaldols occurs via coordination of an oxygen atom of the nitro group with a tin atom, which facilitates hydride delivery in the transition state. The elimination of water from the nitroaldols in mild base is likely driven by the stability of the conjugated nitroalkene products. The elimination required workup with 2N HCl, which likely displaces a nitroalkane-nitroalkene equilibrium towards the latter. These extensions of the Henry reaction lead to products not easily obtained otherwise.

The circadian oscillator gene GIGANTEA mediates a long-term response of the Arabidopsis thaliana circadian clock to sucrose.

Circadian clocks are 24-h timing devices that phase cellular responses; coordinate growth, physiology, and metabolism; and anticipate the day-night cycle. Here we report sensitivity of the Arabidopsis thaliana circadian oscillator to sucrose, providing evidence that plant metabolism can regulate circadian function. We found that the Arabidopsis circadian system is particularly sensitive to sucrose in the dark. These data suggest that there is a feedback between the molecular components that comprise the circadian oscillator and plant metabolism, with the circadian clock both regulating and being regulated by metabolism. We used also simulations within a three-loop mathematical model of the Arabidopsis circadian oscillator to identify components of the circadian clock sensitive to sucrose. The mathematical studies identified GIGANTEA (GI) as being associated with sucrose sensing. Experimental validation of this prediction demonstrated that GI is required for the full response of the circadian clock to sucrose. We demonstrate that GI acts as part of the sucrose-signaling network and propose this role permits metabolic input into circadian timing in Arabidopsis.

Reseña bibliográfica : Rodrik, Dani (2011) The globalization paradox. Democracy and the future of the world economy. W. W. Norton & Co. New York, USA

Dani Rodrik es un conocido economista, nacido en Turquía, Profesor de Economía Política Internacional en la John F. Kennedy School of Government, de Harvard University. Su trabajo se ha destacando en temas de economía internacional, desarrollo económico y economía política. Su área central de investigación se centra en la explicación de que constituye una buena política económica y por qué algunos gobiernos son mejores que otros al adoptar distintas políticas, logrando distintos resultados en materia de desarrollo económico. En este sentido, es un defensor de la estrategia “productivista” del Este de Asia, asignando al Estado y a determinada política económica activa un rol central.

Application of the strain energy density criterion to ductile fracture

The strain energy density criterion due to Sih is used to predict fracture loads of two thin plates subjected to large elastic-plastic deformation. The prediction is achieved with a finite element analysis which is based on Hill's variational principle for incremental deformations capable of solving gross yielding problems involving arbitrary amounts of deformation. The computed results are in excellent agreement with those obtained in Sih's earlier analysis and with an experimental observation.

Congrid eels of the Eastern Pacific and key to their leptocephali

This study indicates that 13 species of congrid larvae belonging to 8 genera occur in the eastern Pacific. The species are: Ariosoma gilberti; Paraconger californiensis; Paraconger sp.; P. dentatus; Chiloconger labiatus; Taenioconger digueti; T. canabus; Gorgasia punctata; G. obtusa; Gnathophis catalinensis; Hildebrandia nitens; Bathycongrus macrurus; and B. varidens. The morphological and anatomical changes undergone during metamorphosis are useful in the identification of the larvae. Larvae are distributed closer to the coastal waters, and are more common from January to May than from June to December. A key to the larvae was developed based on the myotomal counts, adult vertebral counts, pigmentation patterns, and the nature of the teeth and tail tip to distinguish the genera and species. This study shows that Garman's unidentified larvae, Atopichthys acus and A. cingulus, are two different larval stages of Ariosoma gilberti, and points out that Atopichthys dentatus and A. obtusus belong to Paraconger and Gorgasia, respectively. (PDF file contains 25 pages.)

Ichthyoplankton survey of the estuarine and inshore waters of the Florida Everglades, May 1971 to February 1972

Quarterly ichthyoplankton sampling was conducted at 16 estuarine and 24 inshore stations along the Florida Everglades from May 1971 to February 1972. The area is one of the most pristine along lhe Florida coast. The survey provided the first comprehensive information on seasonal occurrence, abundance (under 10 m' of surface area), and distribution of fish eggs and larvae in this area. A total of 209,462 fish eggs and 78,865 larvae was collected. Eggs were identified only as fish eggs, but among the larvae, 37 families, 47 genera, and 37 species were identified. Abundance of eggs and larvae, and diversity of larvae, were greatest in the inshore zone. The 10 most abundant fish families which together made up 90.7% of all larvae from the study area were, in descending order of abundance: Clupeidae, Engraulidae, Gobiidae, Sciaenidae, Carangidae, Pomadasyidae, Cynoglossidae, Gerreidae, Triglidae, and Soleidae. Clupeidae, Engraulidae, and Gobiidae made up 59.9% of all larvae. The inshore zone (to a depth of about 10 m) was a spawning ground and nursery for many fishes important to fisheries. The catch of small larvae (<>3.5 mm SL) indicated that most fishes identified from the 10 most abundant families spawned throughout the inshore zone at depths of <> 10 m, but Orthopristis chrysoptera, Gerreidae, and Prionotus spp. spawned at depths > 10 m, with offshore to inshore (eastward) larval transport. Salinity was one of several environmental factors that probably limited the numbers of eggs and larvae in the estuarine zone. Abundance of eggs and larvae at inshore stations was usually as great as, and sometimes greater than, the abundance of eggs and larvae at offshore stations (due west of the Everglades). (PDF file contains 81 pages.)

Growth, mortality, and exploitation of the Engraulidae, with special reference to the anchoveta, Cetengraulis mysticetus, and the colorado, Anchoa naso, in the Eastern Pacific Ocean

ENGLISH: Growth and mortality data for Cetengraulis mysticetus, Anchoa naso, Engraulis mordax, E. ring ens, E. anchoita, E. encraslcbolus, E. japonicus, and E. australis were assembled and compared. Estimates of the coefficients of natural mortality, M, of E. anchoita and Ancboa naso were made from the maximum age of the former and from data for the other species. The relative yields per recruit at different fishing mortality rates and lengths at entry into the fishery were calculated for each species, using what are considered to be the best estimates and other likely values of K, a constant of growth, and M. The maximum yields per recruit are theoretically obtainable at very high fishing mortality rates, except when the length at entry is low relative to the asymptotic length. K and M may be positively related to the temperature and to each other, and if such is the case at higher temperatures greater fishing effort would be needed to attain the maximum yield per recruit. The applicability of the yield-per-recruit approach to the data is discussed, and suggestions for further research are made. SPANISH: Se reunieron y compararon los datos sobre el crecimiento y mortalidad correspondientes a Cetengraulis mysticetus, Anchoa naso, Engraulis mordax, E. ringens, E. anchoíta, E. encrasicbolus, E. japonicus y E. australls. Los estimativos de los coeficientes de la mortalidad natural, M, de E. anchoita y Anchoa naso se obtuvieron según la edad máxima de E. anchoita y según los datos de las otras especies. Se calculó para cada especie el rendimiento relativo por recluta a diferentes tasas de mortalidad por la pesca y a diferentes longitudes de entrada a la pesquería, empleándose lo que se considera que son los mejores estimativos y otros valores probables de K, una constante de crecímíento, y M. El rendimiento máximo por recluta se obtiene teóricamente a tasas muy altas de la mortalidad por la pesca con excepción de cuando la longitud a la entrada es baja en relación a la longitud asintótica. K y M pueden estar relacionadas positivamente a la temperatura y mutuamente, y si este es el caso a temperaturas más altas se necesitará un esfuerzo superior de pesca para obtener el rendimiento máximo por recluta. La aplicabilidad del enfoque a los datos rendimiento-por-recluta es discutido y se hacen sugerencias para otras investigaciones. (PDF contains 66 pages.)

An extension of the two dimensional JKR theory to the case with a large contact width

In the Hertz and JKR theories, parabolic assumptions for the rounded profiles of the sphere or cylinder are adopted under the condition that the contact radius (width) should be very small compared to the radius of the sphere or cylinder. However, a large contact radius (width) is often found in experiments even under a zero external loading. We aim at extending the plane strain JKR theory to the case with a large contact width. The relation between the external loading and the contact width is given. Solutions for the Hertz, JKR and rounded-profile cases are compared and analyzed. It is found that when the ratio of a/R is approximately larger than about 0.4, the parabolic assumptions in the Hertz and JKR theories are no longer valid and the exact rounded profile function should be used.

Some aspects of the ecology of the limnoplankton, with special reference to the phytoplankton. [Translation from: Svensk Botanisk Tidskrift 13(2) 129-163, 1919.]

This paper tries to develop more generally some fundamental bases for the ecological study of freshwater plankton. A special attention is given to the phytoplankton associations which can be separated out and made into groups according to their dependence upon changing environments. Plankton formations in different types of water bodies (ponds, lakes and rivers) are studied.

I. The 3.7 Å crystal structure of horse heart ferricytochrome C. II. The application of the Karle-Hauptman tangent formula to protein phasing

I. The 3.7 Å Crystal Structure of Horse Heart Ferricytochrome C.

The crystal structure of horse heart ferricytochrome c has been determined to a resolution of 3.7 Å using the multiple isomorphous replacement technique. Two isomorphous derivatives were used in the analysis, leading to a map with a mean figure of merit of 0.458. The quality of the resulting map was extremely high, even though the derivative data did not appear to be of high quality.

Although it was impossible to fit the known amino acid sequence to the calculated structure in an unambiguous way, many important features of the molecule could still be determined from the 3.7 Å electron density map. Among these was the fact that cytochrome c contains little or no α-helix. The polypeptide chain appears to be wound about the heme group in such a way as to form a loosely packed hydrophobic core in the molecule.

The heme group is located in a cleft on the molecule with one edge exposed to the solvent. The fifth coordinating ligand is His 18 and the sixth coordinating ligand is probably neither His 26 nor His 33.

The high resolution analysis of cytochrome c is now in progress and should be completed within the next year.

II. The Application of the Karle-Hauptman Tangent Formula to Protein Phasing.

The Karle-Hauptman tangent formula has been shown to be applicable to the refinement of previously determined protein phases. Tests were made with both the cytochrome c data from Part I and a theoretical structure based on the myoglobin molecule. The refinement process was found to be highly dependent upon the manner in which the tangent formula was applied. Iterative procedures did not work well, at least at low resolution.

The tangent formula worked very well in selecting the true phase from the two possible phase choices resulting from a single isomorphous replacement phase analysis. The only restriction on this application is that the heavy atoms form a non-centric cluster in the unit cell.

Pages 156 through 284 in this Thesis consist of previously published papers relating to the above two sections. References to these papers can be found on page 155.