Genomic analysis of the TRIM family reveals two groups of genes with distinct evolutionary properties

<p>Abstract</p> <p>Background</p> <p>The TRIM family is composed of multi-domain proteins that display the Tripartite Motif (RING, B-box and Coiled-coil) that can be associated with a C-terminal domain. TRIM genes are involved in ubiquitylation and are implicated in a variety of human pathologies, from Mendelian inherited disorders to cancer, and are also involved in cellular response to viral infection.</p> <p>Results</p> <p>Here we defined the entire human TRIM family and also identified the TRIM sets of other vertebrate (mouse, rat, dog, cow, chicken, tetraodon, and zebrafish) and invertebrate species (fruitfly, worm, and ciona). By means of comparative analyses we found that, after assembly of the tripartite motif in an early metazoan ancestor, few types of C-terminal domains have been associated with this module during evolution and that an important increase in TRIM number occurred in vertebrate species concomitantly with the addition of the SPRY domain. We showed that the human TRIM family is split into two groups that differ in domain structure, genomic organization and evolutionary properties. Group 1 members present a variety of C-terminal domains, are highly conserved among vertebrate species, and are represented in invertebrates. Conversely, group 2 is absent in invertebrates, is characterized by the presence of a C-terminal SPRY domain and presents unique sets of genes in each mammal examined. The generation of independent sets of group 2 genes is also evident in the other vertebrate species. Comparing the murine and human TRIM sets, we found that group 1 and 2 genes evolve at different speeds and are subject to different selective pressures.</p> <p>Conclusion</p> <p>We found that the TRIM family is composed of two groups of genes with distinct evolutionary properties. Group 2 is younger, highly dynamic, and might act as a <it>reservoir </it>to develop novel TRIM functions. Since some group 2 genes are implicated in innate immune response, their evolutionary features may account for species-specific battles against viral infection.</p

Family resilience: perception of family members of psychoactive substance dependents

Objective: to understand the perception of family members of psychoactive substance dependents on the elements of the functioning of their family in family resilience. Method: a qualitative approach study, based on the theoretical interpretive framework of family resilience from a systemic perspective. The participants were eleven family members of psychoactive substance dependents from a Psychosocial Care Center - Alcohol and Drugs, from a city in the state of São Paulo. For data collection, semi-structured interview, genogram and ecomap were used. Data analysis was based on the Content Analysis technique, thematic category. Results: from the interviews, three thematic categories were formulated: mobilization in search of support and social support; positive perspectives that would strengthen the family, and assertive communication. These categories point to references to the mobilization and unity of the family in search of social support in the intra-family, extended family and extra-family contexts and positive perspectives, such as persistence, perseverance, hope, faith and religiousness. Conclusion: the situation of having a psychoactive substance dependent in the family seemed to mobilize coping devices and attempts to overcome them through the resilience forces. The results may favor the daily clinical reasoning of the health professionals, helping them to recognize and value the identified resilience attributes.Objetivo: compreender a percepção dos familiares de dependentes de substâncias psicoativas sobre os elementos do funcionamento de sua família na resiliência familiar. Método: estudo de abordagem qualitativa, fundamentado no referencial teórico interpretativo de resiliência familiar na perspectiva sistêmica. Participaram onze familiares de dependentes de substâncias psicoativas de um Centro de Atenção Psicossocial - Álcool e Drogas, de uma cidade do interior paulista. Para a coleta de dados, utilizou-se entrevista semiestruturada, genograma e ecomapa. A análise dos dados baseou-se na técnica de Análise de Conteúdo, categoria temática. Resultados: a partir das entrevistas, foram formuladas três categorias temáticas: mobilização em busca de apoio e suporte social; perspectivas positivas que fortaleceriam a família, e comunicação assertiva. Essas categorias apontam referências à mobilização e união da família em busca de suporte social nos contextos intrafamiliar, de família ampliada e extrafamiliar e perspectivas positivas, como persistência, perseverança, esperança, fé e religiosidade. Conclusão: a situação de ter um dependente de substâncias psicoativas na família pareceu mobilizar dispositivos de enfrentamento e tentativa de superação por intermédio das forças de resiliência. Os resultados podem favorecer o raciocínio clínico cotidiano dos profissionais de saúde, ajudando-os no reconhecimento e na valorização dos atributos de resiliência identificados.Objetivo: comprender la percepción de los familiares de dependientes de sustancias psicoactivas sobre los elementos del funcionamiento de su familia en la resiliencia familiar. Método: estudio cualitativo, basado en el marco teórico de la resiliencia familiar en la perspectiva sistémica. Participaron once familiares de dependientes de sustancias psicoactivas de un Centro de Atención Psicosocial - Alcohol y Drogas, de una ciudad del interior de San Pablo. Para la recolección de datos se utilizaron entrevista semiestructurada, genograma y ecomapa. El análisis de datos se basó en la técnica de Análisis de Contenido, categoría temática. Resultados: a partir de las entrevistas, se formularon tres categorías temáticas: movilización en busca de apoyo y soporte social; perspectivas positivas que fortalezcan a la familia y comunicación asertiva. Estas categorías hacen referencia a la movilización y unidad de la familia en busca de apoyo social en los contextos intrafamiliares, la familia extendida y extrafamiliar y perspectivas positivas, como persistencia, perseverancia, esperanza, fe y religiosidad. Conclusión: tener un dependiente de sustancias psicoactivas en la familia pareció movilizar dispositivos para afrontar e intentar de superar la situación a través de la fuerza de la resiliencia. Los resultados pueden favorecer el razonamiento clínico diario de los profesionales de la salud, ayudándoles a reconocer y valorar los atributos de resiliencia identificados

The James Webb Space Telescope Mission

Twenty-six years ago a small committee report, building on earlier studies, expounded a compelling and poetic vision for the future of astronomy, calling for an infrared-optimized space telescope with an aperture of at least $4m$. With the support of their governments in the US, Europe, and Canada, 20,000 people realized that vision as the $6.5m$ James Webb Space Telescope. A generation of astronomers will celebrate their accomplishments for the life of the mission, potentially as long as 20 years, and beyond. This report and the scientific discoveries that follow are extended thank-you notes to the 20,000 team members. The telescope is working perfectly, with much better image quality than expected. In this and accompanying papers, we give a brief history, describe the observatory, outline its objectives and current observing program, and discuss the inventions and people who made it possible. We cite detailed reports on the design and the measured performance on orbit.Comment: Accepted by PASP for the special issue on The James Webb Space Telescope Overview, 29 pages, 4 figure

MID1 mutations in patients with X-linked Opitz G/BBB syndrome

Mutations in the MID1 gene are responsible for the X-linked form of Opitz G/BBB syndrome (OS), a disorder that affects the development of midline structures. OS is characterized by hypertelorism, hypospadias, laryngo-tracheo-esophageal (LTE) abnormalities, and additional midline defects. Cardiac, anal, and neurological defects are also present. The expressivity of OS is highly variable, even within the same family. We reviewed all the MID1 mutations reported so far, in both familial and sporadic cases. The mutations are scattered along the entire length of the gene and consist of missense and nonsense mutations, insertions and deletions, either in-frame or causing frameshifts, and deletions of either single exons or the entire MID1 coding region. The variety of described mutations and the lack of a strict genotype-phenotype correlation confirm the previous suggestion of the OS phenotype being caused by a loss-of-function mechanism. However, although a specific mutation cannot entirely account for the observed phenotype, we observed preferential association between some types of mutation and specific clinical manifestations, e.g., brain anatomical defects and truncating mutations. This may suggest that the pathogenetic mechanism underlying the OS phenotype is more complex and may vary among the affected organs

Mig12, a novel Opitz syndrome gene product partner, is expressed in the embryonic ventral midline and co-operates with Mid1 to bundle and stabilize microtubules

Opitz G/BBB syndrome is a genetic disorder characterized by developmental midline abnormalities, such as hypertelorism, cleft palate, and hypospadias. The gene responsible for the X-linked form of this disease, MID1, encodes a TRIM/RBCC protein that is anchored to the microtubules. The association of Mid1 with the cytoskeleton is regulated by dynamic phosphorylation, through the interaction with the alpha4 subunit of phosphatase 2A (PP2A). Mid1 acts as an E3 ubiquitin ligase, regulating PP2A degradation on microtubules. In spite of these findings, the biological role exerted by the Opitz syndrome gene product is still unclear and the presence of other potential interacting moieties in the Mid1 structure prompted us to search for additional cellular partners. Through a yeast two-hybrid screening approach, we identified a novel gene, MIG12, whose protein product interacts with Mid1. We confirmed by immunoprecipitation that this interaction occurs in vivo and that it is mediated by the Mid1 coiled-coil domain. We found that Mig12 is mainly expressed in the neuroepithelial midline, urogenital apparatus, and digits during embryonic development. Transiently expressed Mig12 is found diffusely in both nucleus and cytoplasm, although it is enriched in the microtubule-organizing center region. Consistently with this, endogenous Mig12 protein is partially detected in the polymerized tubulin fraction after microtubule stabilization. When co-transfected with Mid1, Mig12 is massively recruited to thick filamentous structures composed of tubulin. These microtubule bundles are resistant to high doses of depolymerizing agents and are composed of acetylated tubulin, thus representing stabilized microtubule arrays. Our findings suggest that Mig12 co-operates with Mid1 to stabilize microtubules. Mid1-Mig12 complexes might be implicated in cellular processes that require microtubule stabilization, such as cell division and migration. Impairment in Mig12/Mid1-mediated microtubule dynamic regulation, during the development of embryonic midline, may cause the pathological signs observed in Opitz syndrome patients

Copper Induction of Laccase Isoenzymes in the Ligninolytic Fungus Pleurotus ostreatus

Pleurotus ostreatus is a white rot basidiomycete that produces several extracellular laccase isoenzymes, including phenol oxidase A1b (POXA1b), POXA2, and POXC. POXC was the most abundant isoenzyme produced under all of the growth conditions examined in this study. Copper was the most efficient inducer of laccase activity among the putative inducers tested. The amounts of all of the previously described laccase isoenzymes increased substantially in copper-supplemented cultures. Under these conditions expression of POX isoenzymes was regulated at the level of gene transcription. It is worth noting that poxa1b mRNA was the most abundant induced transcript at all of the growth times analyzed, and the amount of this mRNA increased until day 7. The discrepancy between the poxa1b transcript and protein amounts can be explained by the presence of a high level of the protein in P. ostreatus cellular extract, which indicated that the POXA1b isoenzyme could be inefficiently secreted and/or that its physiological activity could occur inside the cell or on the cell wall. Moreover, the POXA1b isoenzyme behaved uniquely, as its activity was maximal on the second day of growth and then decreased. An analysis performed with protease inhibitors revealed that the loss of extracellular POXA1b activity could have been due to the presence of specific proteases secreted into the copper-containing culture medium that affected the extracellular POXA1b isoenzyme

Manganese Peroxidase Isoenzymes Produced by Pleurotus ostreatus Grown on Wood Sawdust

The white rot basidiomycete Pleurotus ostreatus produces two manganese peroxidase (MnP) isoenzymes when grown in solid stationary conditions on poplar sawdust, whereas a lower production of these same enzymes is observed on fir sawdust. Addition of Mn21 to poplar culture resulted in a threefold increase of MnP activity; the same addition to fir culture was able to increase tenfold the MnP production. The two MnP isoenzymes (MnP2 and MnP3) were purified from P. ostreatus poplar culture. The isoenzymes differ in their pI values, molecular masses, and N-terminal sequences. MnP3 has the same N-terminal sequence as that of a P. ostreatus MnP previously reported. Both isoenzymes exhibit Mn21-dependent and Mn21-independent peroxidase activities when tested on phenolic substrates. The gene coding for the new isoenzyme MnP2 was cloned and sequenced and the promoter region analyzed. Furthermore, the chromosomal localization of all known P. ostreatus genes was determined

Manganese peroxidase isoenzymes produced by Pleurotus ostreatus grown on wood sawdust.

The white rot basidiomycete Pleurotus ostreatus produces two manganese peroxidase (MnP) isoenzymes when grown in solid stationary conditions on poplar sawdust, whereas a lower production of these same enzymes is observed on fir sawdust. Addition of Mn21 to poplar culture resulted in a threefold increase of MnP activity; the same addition to fir culture was able to increase tenfold the MnP production. The two MnP isoenzymes (MnP2 and MnP3) were purified from P. ostreatus poplar culture. The isoenzymes differ in their pI values, molecular masses, and N-terminal sequences. MnP3 has the same N-terminal sequence as that of a P. ostreatus MnP previously reported. Both isoenzymes exhibit Mn21-dependent and Mn21-independent peroxidase activities when tested on phenolic substrates. The gene coding for the new isoenzyme MnP2 was cloned and sequenced and the promoter region analyzed. Furthermore, the chromosomal localization of all known P. ostreatus genes was determined

Chemical Proteomics Reveals Bolinaquinone as a Clathrin-Mediated Endocytosis Inhibitor

Linking bioactive compounds to their cellular targets is a central challenge in chemical biology. Herein we report the mode of action of perthamide C, a natural cyclopeptide isolated from the marine sponge Theonella swinhoei. Through an emerging mass spectrometry-based chemical proteomics approach, Heat Shock Protein 90 and Glucose Regulated Protein 94 were identified as key targets of perthamide C and this evidence has been validated using surface plasmon resonance. The ability of perthamide C to influence heat shock protein-mediated cell apoptosis revealed that this marine metabolite could be a good candidate for the development of a lead compound with therapeutic applications based on apoptosis modulation