Perturbation of the Nucleus: A Novel Hog1p-independent, Pkc1p-dependent Consequence of Hypertonic Shock in Yeast

Hypertonic shock of Saccharomyces cerevisiae activates the Hog1p MAP kinase cascade. In contrast, protein kinase C (Pkc1p) and the “cell integrity” MAP kinase cascade are critical for the response to hypotonic shock. We observed that hypertonic shock transiently relocated many, but not all, nuclear and nucleolar proteins to the cytoplasm. We hypothesized that the relocation of nuclear proteins was due to activation of the Hog1p kinase cascade, yet, surprisingly, Hog1p was not required for these effects. In contrast, Pkc1p kinase activity was required, although the Pkc1p MAP kinase cascade and several factors known to lie upstream and downstream of Pkc1p were not. Moreover, sudden induction of a hyperactive form of Pkc1p was sufficient to relocate nuclear proteins. Taken together, these observations show that the scope of involvement of Pkc1p in the organization of the nucleus considerably exceeds what has been characterized previously. The relocation of nuclear proteins is likely to account for the profound inhibition of RNA synthesis that was observed during hypertonic shock.

Operator binding by lambda repressor heterodimers with one or two N-terminal arms.

The first 6 amino acids (NH2-Ser1-Thr2-Lys3-Lys4-Lys5-Pro6) of bacteriophage lambda cI repressor form a flexible arm that wraps around the operator DNA. Homodimeric lambda repressor has two arms. To determine whether both arms are necessary or only one arm is sufficient for operator binding, we constructed heterodimeric repressors with two, one, or no arms by fusing the DNA binding domain of lambda repressor to leucine zippers from Fos and Jun. Although only one arm is visible in the cocrystal structure of the N-domain-operator complex, our results indicate that both arms are required for optimal operator binding and normal site discrimination.

Binding of purified multiple antibiotic-resistance repressor protein (MarR) to mar operator sequences.

Elevated expression of the marORAB multiple antibiotic-resistance operon enhances the resistance of Escherichia coli to various medically significant antibiotics. Transcription of the operon is repressed in vivo by the marR-encoded protein, MarR, and derepressed by salicylate and certain antibiotics. The possibility that repression results from MarR interacting with the marO operator-promoter region was studied in vitro using purified MarR and a DNA fragment containing marO. MarR formed at least two complexes with marO DNA, bound > 30-fold more tightly to it than to salmon sperm DNA, and protected two separate 21-bp sites within marO from digestion by DNase I. Site I abuts the downstream side of the putative -35 transcription-start signal and includes 4 bp of the -10 signal. Site II begins 13 bp downstream of site I, ending immediately before the first base pair of marR. Site II, approximately 80% homologous to site I, is not required for repression since a site II-deleted mutant (marO133) was repressed in trans by wild-type MarR. The absence of site II did not prevent MarR from complexing with the site I of marO133. Salicylate bound to MarR (Kd approximately 0.5 mM) and weakened the interaction of MarR with sites I and II. Thus, repression of the mar operon, which curbs the antibiotic resistance of E. coli, correlates with the formation of MarR-site I complexes. Salicylate appears to induce the mar operon by binding to MarR and inhibiting complex formation, whereas tetracycline and chloramphenicol, which neither bind MarR nor inhibit complex formation, must induce by an indirect mechanism.

Formação do preço da energia convencional nas transações entre agentes no mercado de curto prazo brasileiro.

Hoje no mercado brasileiro de eletricidade, o preço da energia convencional é composto pela soma do valor do Preço de Liquidação das Diferenças (PLD) divulgado pela Câmara de Comercialização de Energia Elétrica (CCEE) semanalmente com o valor do Spread negociado bilateralmente no mercado à vista (mercado de curto prazo), resultante do equilíbrio entre oferta e demanda. Em alguns momentos, o valor do Spread chega a representar mais de 100% do custo total da energia. Este trabalho faz uma análise do mercado brasileiro, bem como, de alguns mercados no exterior de energia elétrica e destaca os pontos que tem influência direta, na formação do Spread da energia convencional e como isso afeta a decisão de contratação dos agentes. Além disso, o trabalho busca encontrar correlações entre dados divulgados, como carga e oferta de energia, com o ágio negociado no mercado de curto prazo, buscando entender o real impacto de cada um desses fatores e explicar as grandes variações já observadas. Sugere-se também um modelo de regressão linear múltipla para a projeção de valores do ágio. Para tanto, foram utilizadas informações proveniente de um banco de dados de cotações de negócios efetivamente realizados no curto prazo desde janeiro de 2011 até julho de 2014, bem como informações retiradas da CCEE e Operador Nacional do Sistema (ONS).

Motorcycle operator licensing - issue paper.

Mode of access: Internet.

Social decision-making for high consequence, low probability occurences /

Oct. 1978.

Results of owner/operator and mechanic/technician survey. Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Recreational boat safe loading : operator study /

Contract no. DOT-CG-40672-A.

Development of a revised version of the NHTSA Motorcycle Operator Skill Test (MOST II). Final report.

National Highway Traffic Safety Administration, Washington, D.C.

Crash test of cone truck for operator safety. Final report.

Mode of access: Internet.

Development of a low-cost truck weighing system design and operator manual. Final report.

Federal Highway Administration, Office of Engineering and Highway Operations Research and Development, McLean, Va.