Characterization of the human SCF ubiquitin ligases - structure, function, and regulation

The SCF ubiquitin ligase complex of budding yeast triggers DNA replication by cata lyzi ng ubiquitination of the S phase CDK inhibitor SIC1. SCF is composed of several evolutionarily conserved proteins, including ySKP1, CDC53 (Cullin), and the F-box protein CDC4. We isolated hSKP1 in a two-hybrid screen with hCUL1, the human homologue of CDC53. We showed that hCUL1 associates with hSKP1 in vivo and directly interacts with hSKP1 and the human F-box protein SKP2 in vitro, forming an SCF-Iike particle. Moreover, hCUL1 complements the growth defect of yeast CDC53^(ts) mutants, associates with ubiquitination-promoting activity in human cell extracts, and can assemble into functional, chimeric ubiquitin ligase complexes with yeast SCF components. These data demonstrated that hCUL1 functions as part of an SCF ubiquitin ligase complex in human cells. However, purified human SCF complexes consisting of CUL1, SKP1, and SKP2 are inactive in vitro, suggesting that additional factors are required.

Subsequently, mammalian SCF ubiquitin ligases were shown to regulate various physiological processes by targeting important cellular regulators, like lĸBα, β-catenin, and p27, for ubiquitin-dependent proteolysis by the 26S proteasome. Little, however, is known about the regulation of various SCF complexes. By using sequential immunoaffinity purification and mass spectrometry, we identified proteins that interact with human SCF components SKP2 and CUL1 in vivo. Among them we identified two additional SCF subunits: HRT1, present in all SCF complexes, and CKS1, that binds to SKP2 and is likely to be a subunit of SCF5^(SKP2) complexes. Subsequent work by others demonstrated that these proteins are essential for SCF activity. We also discovered that COP9 Signalosome (CSN), previously described in plants as a suppressor of photomorphogenesis, associates with CUL1 and other SCF subunits in vivo. This interaction is evolutionarily conserved and is also observed with other Cullins, suggesting that all Cullin based ubiquitin ligases are regulated by CSN. CSN regulates Cullin Neddylation presumably through CSNS/JAB1, a stochiometric Signalosome subunit and a putative deneddylating enzyme. This work sheds light onto an intricate connection that exists between signal transduction pathways and protein degradation machinery inside the cell and sets stage for gaining further insights into regulation of protein degradation.

Descripción de la morfología externa e interna del langostino con algunas aplicaciones de indole taxonómica y biológica

In the present paper the external and internal morphology of the shrimp Pleoticus muelleri (Bate), is described in detail. Special attention is given to those features which are of diagnostic value or which are significant for a fuller understanding of the biology of this species. The major portion of the paper deals with the description of the cephalopereion, pleon and telson and its appendages, as well as the nervous, respiratory, circulatory, digestive, excretory and reproductive systems. Special attention is given to the digestive system, the stomodaeal apparatus and sexual dimorphism. The relations between form and function of several organs and systems are pointed out. A study is made of the development of the gonads, the sexual cycle in males and females; the variations found in relative weight and size are described. The phylogenetical jposition of the Penaeidae, the genera of the subfamily Solenoceridae and of P. muelleri are discussed. (PDF contains 72 pages)

Edge function method applied to thin plates resting on Winkler foundation

The Edge Function method formerly developed by Quinlan⁽²⁵⁾ is applied to solve the problem of thin elastic plates resting on spring supported foundations subjected to lateral loads the method can be applied to plates of any convex polygonal shapes, however, since most plates are rectangular in shape, this specific class is investigated in this thesis. The method discussed can also be applied easily to other kinds of foundation models (e.g. springs connected to each other by a membrane) as long as the resulting differential equation is linear. In chapter VII, solution of a specific problem is compared with a known solution from literature. In chapter VIII, further comparisons are given. The problems of concentrated load on an edge and later on a corner of a plate as long as they are far away from other boundaries are also given in the chapter and generalized to other loading intensities and/or plates springs constants for Poisson's ratio equal to 0.2

The gated integration technique for the accurate measurement of the autocorrelation function of speckle intensities scattered from random phase screens

A new approach based on the gated integration technique is proposed for the accurate measurement of the autocorrelation function of speckle intensities scattered from a random phase screen. The Boxcar used for this technique in the acquisition of the speckle intensity data integrates the photoelectric signal during its sampling gate open, and it repeats the sampling by a preset number, in. The average analog of the in samplings output by the Boxcar enhances the signal-to-noise ratio by root m, because the repeated sampling and the average make the useful speckle signals stable, while the randomly varied photoelectric noise is suppressed by 1/ root m. In the experiment, we use an analog-to-digital converter module to synchronize all the actions such as the stepped movement of the phase screen, the repeated sampling, the readout of the averaged output of the Boxcar, etc. The experimental results show that speckle signals are better recovered from contaminated signals, and the autocorrelation function with the secondary maximum is obtained, indicating that the accuracy of the measurement of the autocorrelation function is greatly improved by the gated integration technique. (C) 2006 Elsevier Ltd. All rights reserved.

Climate change impacts on the water services in Costa Rica: a production function for the hydroenergy sector

23 p.

Prevalência de HPV em adolescentes virgens e com atividade sexual

O Papilomavírus humano (HPV) é um vírus DNA bastante prevalente em estimativas mundiais, sendo a DST isolada mais frequente no mundo. Existem poucos dados sobre prevalência em adolescentes sem vida sexual ativa, havendo ainda muitos tabus nesta faixa etária. O presente estudo tem por objetivo identificar a prevalência de Papilomavírus humano através de biologia molecular em pacientes sem coitarca, comparando com um grupo de pacientes da mesma faixa etária com atividade sexual. Foram avaliadas 100 adolescentes com idade variando de 11 a 20 anos, com pelo menos dois anos pós-menarca, atendidas no período de janeiro de 2007 a janeiro de 2009 no ambulatório de ginecologia infantopuberal do Hospital Universitário Pedro Ernesto. Das 100 adolescentes, 50 apresentavam hímen íntegro e 50 relatavam atividade sexual regular. Para as pacientes sem coitarca (grupo 1) foi realizada uma coleta apenas de vestíbulo e para as pacientes com atividade sexual (grupo 2) foi realizada coleta de vagina e endocérvice. A pesquisa de DNA-HPV foi realizada por captura híbrida de 2 geração. Os resultados foram descritos em unidade relativa de luz. Para os dados categóricos foram aplicados os testes exato de Fisher. A positividade geral de DNA de HPV de alto e de baixo risco foi de 72%. No grupo 1 houve positividade do teste em 3 casos (6%). Já no grupo 2, 33 casos (66%) foram positivos para pelo menos um sítio. Material de vagina foi positivo em 30 casos (60%) e de colo em 27 casos (54%). Pode-se concluir que a positividade nas meninas com vida sexual ativa é alta. A infecção pelo HPV, ainda que pouco frequente, pode estar presente em meninas sem coitarca.

Sequence-function relationships in E. coli transcriptional regulation

Understanding how transcriptional regulatory sequence maps to regulatory function remains a difficult problem in regulatory biology. Given a particular DNA sequence for a bacterial promoter region, we would like to be able to say which transcription factors bind there, how strongly they bind, and whether they interact with each other and/or RNA polymerase, with the ultimate objective of integrating knowledge of these parameters into a prediction of gene expression levels. The theoretical framework of statistical thermodynamics provides a useful framework for doing so, enabling us to predict how gene expression levels depend on transcription factor binding energies and concentrations. We used thermodynamic models, coupled with models of the sequence-dependent binding energies of transcription factors and RNAP, to construct a genotype to phenotype map for the level of repression exhibited by the lac promoter, and tested it experimentally using a set of promoter variants from E. coli strains isolated from different natural environments. For this work, we sought to ``reverse engineer'' naturally occurring promoter sequences to understand how variations in promoter sequence affects gene expression. The natural inverse of this approach is to ``forward engineer'' promoter sequences to obtain targeted levels of gene expression. We used a high precision model of RNAP-DNA sequence dependent binding energy, coupled with a thermodynamic model relating binding energy to gene expression, to predictively design and verify a suite of synthetic E. coli promoters whose expression varied over nearly three orders of magnitude.

However, although thermodynamic models enable predictions of mean levels of gene expression, it has become evident that cell-to-cell variability or ``noise'' in gene expression can also play a biologically important role. In order to address this aspect of gene regulation, we developed models based on the chemical master equation framework and used them to explore the noise properties of a number of common E. coli regulatory motifs; these properties included the dependence of the noise on parameters such as transcription factor binding strength and copy number. We then performed experiments in which these parameters were systematically varied and measured the level of variability using mRNA FISH. The results showed a clear dependence of the noise on these parameters, in accord with model predictions.

Finally, one shortcoming of the preceding modeling frameworks is that their applicability is largely limited to systems that are already well-characterized, such as the lac promoter. Motivated by this fact, we used a high throughput promoter mutagenesis assay called Sort-Seq to explore the completely uncharacterized transcriptional regulatory DNA of the E. coli mechanosensitive channel of large conductance (MscL). We identified several candidate transcription factor binding sites, and work is continuing to identify the associated proteins.

Measurement of the neutron (^3He) spin structure function at low Q^2 : a connection between the Bjorken and Drell-Hearn-Gerasimov sum rules

The spin dependent cross sections, σT_1/2 and σT_3/2 , and asymmetries, A_∥ and A_⊥ for ³He have been measured at the Jefferson Lab's Hall A facility. The inclusive scattering process ³He(e,e)X was performed for initial beam energies ranging from 0.86 to 5.1 GeV, at a scattering angle of 15.5°. Data includes measurements from the quasielastic peak, resonance region, and the deep inelastic regime. An approximation for the extended Gerasimov-Drell-Hearn integral is presented at a 4-momentum transfer Q² of 0.2-1.0 GeV².

Also presented are results on the performance of the polarized ³He target. Polarization of ³He was achieved by the process of spin-exchange collisions with optically pumped rubidium vapor. The ³He polarization was monitored using the NMR technique of adiabatic fast passage (AFP). The average target polarization was approximately 35% and was determined to have a systematic uncertainty of roughly ±4% relative.

O tema do excesso sexual em Gilberto Freyre : Casa Grande & Senzala e Sobrados e Mucambos

Esta dissertação tem por objeto a imagem da sexualidade excessiva do brasileiro, como um dos elementos de caracterização da identidade nacional. Toma as obras CasaGrande & Senzala, e Sobrados e Mocambos de Gilberto Freyre como centrais para a difusão dessa crença. A narrativa freyriana combina de forma já observada por diversos críticos, uma alternância entre ênfases mais naturalizantes e outras enraizadas na esfera cultural. No primeiro caso, por exemplo, estabelece nexos entre raça, sexo e clima. No segundo caso, valoriza o papel da escravidão para caracterizar aspectos da miscigenação e da sexualidade, presentes na sociedade brasileira. Salienta-se o modo como o autor construiu seu discurso sobre as relações entre homens e mulheres, negros, índios, mulatos e brancos. Estas são fundadas em categorias opositivas que revelam uma constante na atribuição de predicados que conectam sexo e gênero, raça e etnia, a partir de um viés assimétrico. Tal procedimento analítico sugere um persistente idioma de gênero.

Environmental oestrogens and sexual development in fish

This brief paper discusses the assumption that watercourses might be harbouring a chemical(s) affecting the sexual development in fish. Male fish was found with the oestrogen-dependent blood protein, vitellogenin, usually found only in maturing females. The author examines a number of man-made chemicals present in the environment have been found to be oestrogenic. The paper concludes that rivers contain environmental oestrogens that are capable of causing disruptions in the sexual development of fish. Whether or not these environmental oestrogens are causing a widespread disruption in reproduction in wild fish, however, has yet to be determined.