A protein kinase screen of Neurospora crassa mutant strains reveals that the SNF1 protein kinase promotes glycogen synthase phosphorylation

Glycogen functions as a carbohydrate reserve in a variety of organisms and its metabolism is highly regulated. The activities of glycogen synthase and glycogen phosphorylase, the rate-limiting enzymes of the synthesis and degradation processes, respectively, are regulated by allosteric modulation and reversible phosphorylation. To identify the protein kinases affecting glycogen metabolism in Neurospora crassa, we performed a screen of 84 serine/threonine kinase knockout strains. We identified multiple kinases that have already been described as controlling glycogen metabolism in different organisms, such as NcSNF1, NcPHO85, NcGSK3, NcPKA, PSK2 homologue and NcATG1. In addition, many hypothetical kinases have been implicated in the control of glycogen metabolism. Two kinases, NcIME-2 and NcNIMA, already functionally characterized but with no functions related to glycogen metabolism regulation, were also identified. Among the kinases identified, it is important to mention the role of NcSNF1. We showed in the present study that this kinase was implicated in glycogen synthase phosphorylation, as demonstrated by the higher levels of glycogen accumulated during growth, along with a higher glycogen synthase (GSN) ±glucose 6-phosphate activity ratio and a lesser set of phosphorylated GSN isoforms in strain Ncsnf1KO, when compared with the wild-type strain. The results led us to conclude that, in N. crassa, this kinase promotes phosphorylation of glycogen synthase either directly or indirectly, which is the opposite of what is described for Saccharomyces cerevisiae. The kinases also play a role in gene expression regulation, in that gdn, the gene encoding the debranching enzyme, was down-regulated by the proteins identified in the screen. Some kinases affected growth and development, suggesting a connection linking glycogen metabolism with cell growth and development.

O Wortmannin reverte a hiporreatividade de aortas à fenilefrina associada à fase final da prenhez de ratas espontaneamente hipertensas (SHR)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resposta individual à ingestão de treonina para suínos em fase de crescimento

Pós-graduação em Zootecnia - FCAV

Modelagem do crescimento, maturação sexual e produção de ovos como ferramenta para estimar as exigências nutricionais de matrizes pesadas

Pós-graduação em Zootecnia - FCAV

Avaliação da fosforilação da Akt e do conteúdo da proteína transportadora de glicose GLUT4 em músculo esquelético de ratos adultos com lesão periapical

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antitumor activity of irradiated riboflavin on human renal carcinoma cell line 786-O

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Site and extent of amino acid digestion in dairy cattle fed with corn and its byproducts

The study was conducted to evaluated the site and extent of dry matter (DM), crude protein (CP), methionine (Met), lysine (Lys), and threonine (Thr) digestion of corn and byproducts obtained from corn germ mixed with different amounts of extruded or non-extruded ether extract (EE) in dairy cattle. Treatments consisted in eight types of feed and two processing in a 4 x 2 factorial design. There were four feeds: corn grain cracked (Corn), corn germ meal with 1% EE (CG1), corn germ meal with 7% EE (CG7), and corn germ meal with 10% EE (CG10). The feeds were processed in one of two ways: extruded (Ex) and not extruded. In situ techniques were used to determine DM, CP, Met, Lys, and Thr partial and total tract digestion. A basic diet was compounded of corn germ meal, soybean meal and coastcross hay in a 70: 30 roughage to concentrate ratio. There was no interaction (P>0.05) between feeds and processing method. Extrusion improved (P<0.05) total tract digestibility of corn DM but not CP. Intestinal digestibility was similar (P>0.05) for corn and corn germ meal mixed with 7 and 10% EE, regardless of EE processing method. The CP total tract digestibility of corn germ meal with 1% non-extruded EE was 16.62% higher (P<0.05) than that of the extruded form. The best total CP digestibility was obtained for corn germ meal with 7% EE, independently of the processing method. The effects of EE processing method on partial and total digestibility differed between amino acid. Corn and corn byproduct extrusion may improve dry matter digestibility, but do not necessarily influence crude protein digestion. Ruminal and intestinal digestibility of Met, Lys, and Thr depends on both feed type and processing method. Therefore, amino acid availability should be considered individually.

Phytotoxicity associated to microcystins: a review

Microcystins (MC) are the most studied toxins of cyanobacteria since they are widely distributed and account for several cases of human and animal poisoning, being potent inhibitors of the serine/threonine protein phosphatases 1 (PP1) and 2A (PP2A). The phosphatases PP1 and PP2A are also present in plants, which may also suffer adverse effects due to the inhibition of these enzymes. In aquatic plants, biomass reduction is usually observed after absorption of cyanotoxins, which can bioaccumulate in its tissues. In terrestrial plants, the effects caused by microcystins vary from inhibition to stimulation as the individuals develop from seedling to adult, and include reduction of protein phosphatases 1 and 2A, oxidative stress, decreased photosynthetic activity and even cell apoptosis, as well as bioaccumulation in plant tissues. Thus, the irrigation of crop plants by water contaminated with microcystins is not only an economic problem but becomes a public health issue because of the possibility of food contamination, and this route of exposure requires careful monitoring by the responsible authorities.

Mycorrhization alters foliar soluble amino acid composition and influences tolerance to Pb in Calopogonium mucunoides

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The Mycobacterium leprae hsp65 displays proteolytic activity. Mutagenesis studies indicate that the M. leprae hsp65 Proteolytic activity is catalytically related to the HslVU protease?

The present study reports, for the first time, that the recombinant hsp65 from Mycobacterium leprae (chaperonin 2) displays a proteolytic activity toward oligopeptides. The M. leprae hsp65 proteolytic activity revealed a trypsin-like specificity toward quenched fluorescence peptides derived from dynorphins. When other peptide substrates were used (β-endorphin, neurotensin, and angiotensin I), the predominant peptide bond cleavages also involved basic amino acids in P 1, although, to a minor extent, the hydrolysis involving hydrophobic and neutral amino acids (G and F) was also observed. The amino acid sequence alignment of the M. leprae hsp65 with Escherichia coli Hs1VU protease suggested two putative threonine catalytic groups, one in the N-domain (T 136, K 168, and Y 264) and the other in the C-domain (T 375, K 409, and S 502). Mutagenesis studies showed that the replacement of K 409 by A caused a complete loss of the proteolytic activity, whereas the mutation of K 168 to A resulted in a 25% loss. These results strongly suggest that the amino acid residues T 375, K 409, and S 502 at the C-domain form the catalytic group that carries out the main proteolytic activity of the M. leprae hsp65. The possible pathophysiological implications of the proteolytic activity of the M. leprae hsp65 are now under investigation in our laboratory.

Produção, caracterização nutricional e utilização de farinhas e óleos de resíduos de peixes neotropicais em dietas para Tilápia do Nilo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Relação ideal dos aminoácidos essenciais para mantença, crescimento e produção de aves

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Eficiência da utilização e turnover do nitrogênio da L-(15N) treonina em tecidos de frango de corte

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeitos da ingestão de simbiótico e indol-3-carbinol sobre o processo de carcinogênese química de cólon em ratos Wistar alimentados com dieta contendo heme

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)