Human PEX1 cloned by functional complementation on a CHO cell mutant is responsible for peroxisome-deficient Zellweger syndrome of complementation group I

The peroxisome biogenesis disorders (PBDs), including Zellweger syndrome (ZS) and neonatal adrenoleukodystrophy (NALD), are autosomal recessive diseases caused by defects in peroxisome assembly, for which at least 10 complementation groups have been reported. We have isolated a human PEX1 cDNA (HsPEX1) by functional complementation of peroxisome deficiency of a mutant Chinese hamster ovary (CHO) cell line, ZP107, transformed with peroxisome targeting signal type 1-tagged “enhanced” green fluorescent protein. This cDNA encodes a hydrophilic protein (Pex1p) comprising 1,283 amino acids, with high homology to the AAA-type ATPase family. A stable transformant of ZP107 with HsPEX1 was morphologically and biochemically restored for peroxisome biogenesis. HsPEX1 expression restored peroxisomal protein import in fibroblasts from three patients with ZS and NALD of complementation group I (CG-I), which is the highest-incidence PBD. A CG-I ZS patient (PBDE-04) possessed compound heterozygous, inactivating mutations: a missense point mutation resulting in Leu-664 → Pro and a deletion of the sequence from Gly-634 to His-690 presumably caused by missplicing (splice site mutation). Both PBDE-04 PEX1 cDNAs were defective in peroxisome-restoring activity when expressed in the patient fibroblasts as well as in ZP107 cells. These results demonstrate that PEX1 is the causative gene for CG-I peroxisomal disorders.

Herpes simplex virus 1 α regulatory protein ICP0 functionally interacts with cellular transcription factor BMAL1

The infected cell protein no. 0 (ICP0) of herpes simplex virus 1 (HSV-1) is a promiscuous transactivator shown to enhance the expression of gene introduced into cells by infection or transfection. At the molecular level, ICP0 is a 775-aa ring finger protein localized initially in the nucleus and late in infection in the cytoplasm and mediates the degradation of several proteins and stabilization of others. None of the known functions at the molecular level account for the apparent activity of ICP0 as a transactivator. Here we report that ICP0 functionally interacts with cellular transcription factor BMAL1, a member of the basic helix–loop–helix PER-ARNT-SIM (PAS) super family of transcriptional regulators. Specifically, sequences mapped to the exon II of ICP0 interacted with BMAL1 in the yeast two-hybrid system and in reciprocal pull-down experiments in vitro. Moreover, the enhancement of transcription of a luciferase reporter construct whose promoter contained multiple BMAL1-binding sites by ICP0 and BMAL1 was significantly greater than that observed by ICP0 or BMAL1 alone. Although the level of BMAL1 present in nuclei of infected cells remained unchanged between 3 and 8 h after infection, the level of cytoplasmic BMAL1 was reduced at 8 h after infection. The reduction of cytoplasmic BMAL1 was significantly greater in cells infected with the ICP0-null mutant than in the wild-type virus-infected cells, suggesting that ICP0 mediates partial stabilization of the protein. These results indicate that ICP0 interacts physically and functionally with at least one cellular transcription-regulatory factor.

Ccm1, a regulatory gene controlling the induction of a carbon-concentrating mechanism in Chlamydomonas reinhardtii by sensing CO2 availability

Aquatic photosynthetic organisms, including the green alga Chlamydomonas reinhardtii, induce a set of genes for a carbon-concentrating mechanism (CCM) to acclimate to CO2-limiting conditions. This acclimation is modulated by some mechanisms in the cell to sense CO2 availability. Previously, a high-CO2-requiring mutant C16 defective in an induction of the CCM was isolated from C. reinhardtii by gene tagging. By using this pleiotropic mutant, we isolated a nuclear regulatory gene, Ccm1, encoding a 699-aa hydrophilic protein with a putative zinc-finger motif in its N-terminal region and a Gln repeat characteristic of transcriptional activators. Introduction of Ccm1 into this mutant restored an active carbon transport through the CCM, development of a pyrenoid structure in the chloroplast, and induction of a set of CCM-related genes. That a 5,128-base Ccm1 transcript and also the translation product of 76 kDa were detected in both high- and low-CO2 conditions suggests that CCM1 might be modified posttranslationally. These data indicate that Ccm1 is essential to control the induction of CCM by sensing CO2 availability in Chlamydomonas cells. In addition, complementation assay and identification of the mutation site of another pleiotropic mutant, cia5, revealed that His-54 within the putative zinc-finger motif of the CCM1 is crucial to its regulatory function.

Resistência ao cisalhamento de contactos solo-rocha

Nos estudos referentes à estabilidade de maciços rochosos, é de capital importância a identificação dos planos de fraqueza do maciço rochoso. Estes planos de fraques condicionam a estabilidade do maciço, tanto mais quanto mais adversa a posição dos mesmos com relação ás tensões cisalhantes atuantes. De modo geral, os planos de fraqueza consistem de camadas e intercalações de rocha fraca ou alterada, ou de fraturas e outras descontinuidades as quais são frequentemente preenchidas por solo. Quando uma intercalação de solo, argilito, folhelho, etc, apresenta-se em intercalação a camadas de rochas duras, (ou mesmo de preenchimentos alterados de fraturas e falhas) tem sido prática comum adotar a resistência ao cisalhamento do material de preenchimento como sendo o valor mais baixo da resistência da descontinuidade. Entretanto, conforme foi sugerido por Patton (1968), existe a hipótese de que a prática acima não seja válida, e que a resistência ao cisalhamento do contacto seja inferior à obtida para o solo de preenchimento propriamente dito. O objetivo desta investigação é o de iniciar o estudo da resistência ao cisalhamento de contactos solo-rocha, e de compara essa resistência às do solo e da rocha obtidas separadamente

El liderazgo en Colombia : un análisis de la investigación empírica en contextos organizacionales

El objetivo de este estudio fue establecer las diferentes tendencias de la investigación sobre el Liderazgo en las organizaciones colombianas, mediante un estudio bibliométrico donde se analizaron 21 artículos publicados en revistas de Publindex, dentro de las categorías a1, a2, b y c. Para el análisis de los artículos se realizó una matriz en Excel que permitió registrar variables como estilos de liderazgo, resultados, años de publicación, revistas y autores. De acuerdo con los resultados se evidenció que el tema ha sido estudiado de forma significativa pero aún son pocas las conclusiones que permiten esclarecer el liderazgo como concepto en Colombia. Sin embargo, hay hallazgos importantes como los estilos de liderazgo predominantes: transformacional y transaccional; se encuentran variables psicológicas asociadas al estudio de liderazgo más comunes como inteligencia emocional, clima organizacional, cultura organizacional y los instrumentos más utilizados: entrevistas y cuestionarios. Se concluye que Colombia es un país que si investiga sobre esta temática, sin embargo, no hay acuerdos y hallazgos unificados. Se espera que en el futuro se continúe promoviendo una co-contrucción del concepto.