Discrete cytosolic macromolecular BRAF complexes exhibit distinct activities and composition

As a central element within the RAS/ERK pathway, the serine/threonine kinase BRAF plays a key role in development and homeostasis and represents the most frequently mutated kinase in tumors. Consequently, it has emerged as an important therapeutic target in various malignancies. Nevertheless, the BRAF activation cycle still raises many mechanistic questions as illustrated by the paradoxical action and side effects of RAF inhibitors. By applying SEC‐PCP‐SILAC, we analyzed protein–protein interactions of hyperactive BRAFV600E and wild‐type BRAF (BRAFWT). We identified two macromolecular, cytosolic BRAF complexes of distinct molecular composition and phosphorylation status. Hyperactive BRAFV600E resides in large complexes of higher molecular mass and activity, while BRAFWT is confined to smaller, slightly less active complexes. However, expression of oncogenic K‐RasG12V, either by itself or in combination with RAF dimer promoting inhibitors, induces the incorporation of BRAFWT into large, active complexes, whereas pharmacological inhibition of BRAFV600E has the opposite effect. Thus, the quaternary structure of BRAF complexes is shaped by its activation status, the conformation of its kinase domain, and clinically relevant inhibitors

A Study of Feminine Role Models Selected by Catholic Women

Women are being challenged to greater levels of growth and self-development because of the expanding opportunities and options available to them in nearly ever facet of society today. Most women need the support and inspiration of other women as role models in order to respond to these challenges. In living out their faith commitment in the present culture, Catholic women also need role models. The Catholic Church has long presented its members with a variety of saints, and with Mary, the mother of Jesus, as role models. Are Mary and women saints viable role models for Catholic women today? If not, who are the women who are serving as role models for them? This descriptive study utilized survey research methods to examine the feminine role models of Catholic women in order to determine the type of women who serve as role models for them, the specific prominent individuals whom they select as their role models, and the qualities select. Participants in the study were women volunteers of various ages, marital states and education, from rural, urban and suburban areas in six different states. Three hundred sixty-two women responded to the survey in which they named and identified women whom they considered role models for themselves. The study concluded that women of all ages do select feminine role models for themselves, and that a woman\u27s own mother is most likely to be among her role models. Married women who are personally known to the individual are selected most frequently. Prominent women, when selected, are likely to be humanitarians such as Mother Teresa of Calcutta or the wives of United States presidents. The qualities most frequently admired in the role models selected are those of loving and caring, courage, service, strength, ability, and faith. Mary, the mother of Jesus, is not widely selected as a role model by Catholic women, and women saints of the Catholic Church are least likely to be selected as role models by Catholic women

Optimizing Nervous System-Specific Gene Targeting with Cre Driver Lines: Prevalence of Germline Recombination and Influencing Factors.

The Cre-loxP system is invaluable for spatial and temporal control of gene knockout, knockin, and reporter expression in the mouse nervous system. However, we report varying probabilities of unexpected germline recombination in distinct Cre driver lines designed for nervous system-specific recombination. Selective maternal or paternal germline recombination is showcased with sample Cre lines. Collated data reveal germline recombination in over half of 64 commonly used Cre driver lines, in most cases with a parental sex bias related to Cre expression in sperm or oocytes. Slight differences among Cre driver lines utilizing common transcriptional control elements affect germline recombination rates. Specific target loci demonstrated differential recombination; thus, reporters are not reliable proxies for another locus of interest. Similar principles apply to other recombinase systems and other genetically targeted organisms. We hereby draw attention to the prevalence of germline recombination and provide guidelines to inform future research for the neuroscience and broader molecular genetics communities

Chd8 mediates cortical neurogenesis via transcriptional regulation of cell cycle and Wnt signaling

De novo mutations in CHD8 are strongly associated with autism spectrum disorder, but the basic biology of CHD8 remains poorly understood. Here we report that Chd8 knockdown during cortical development results in defective neural progenitor proliferation and differentiation that ultimately manifests in abnormal neuronal morphology and behaviors in adult mice. Transcriptome analysis revealed that while Chd8 stimulates the transcription of cell cycle genes, it also precludes the induction of neural-specific genes by regulating the expression of PRC2 complex components. Furthermore, knockdown of Chd8 disrupts the expression of key transducers of Wnt signaling, and enhancing Wnt signaling rescues the transcriptional and behavioral deficits caused by Chd8 knockdown. We propose that these roles of Chd8 and the dynamics of Chd8 expression during development help negotiate the fine balance between neural progenitor proliferation and differentiation. Together, these observations provide new insights into the neurodevelopmental role of Chd8.National Institutes of Health (U.S.) (Grant UH1-MH106018-03

The RAB3-RIM Pathway Is Essential for the Release of Neuromodulators

Neurons secrete neuromodulators/neuropeptides from dense-core vesicles (DCVs) by a largely unknown mechanism. Persoon et al. identify RAB3 and RIM1/2 as essential factors. RAB3’s indispensable role is the first distinct feature of DCV secretion as compared to synaptic vesicle secretion

Utilização de marcadores moleculares para identificação de híbridos e associação com apomixia em Brachiaria.

O gênero Brachiaria contem diversas espécies forrageiras amplamente utilizadas na pecuária de leite e carne em regiões de clima tropical. A grande maioria das pastagens cultivadas no Brasil pertence ao gênero Brachiaria o que reforça a enorme importância econômica do gênero. A Embrapa coordena programas de melhoramento em Brachiaria visando a produção de materiais mais produtivos e resistente à doenças e pragas. Os programas de melhoramento utilizam diferentes estratégias para a seleção de materiais superiores e uma das estratégias consiste na formação de híbridos interespecíficos. A confirmação da hibridação é um dos problemas existentes e ferramentas moleculares podem ser utilizadas para auxiliar nesse processo de identificação. Outro fator que também pode ser auxiliado pelas ferramentas moleculares é a identificação de materiais apomíticos, característica muito importante e que pode direcionar estratégias completamente diferentes dentro de um programa de melhoramento. Este trabalho buscou identificar híbridos entre B. brizantha e B. ruziziensis em diferentes cruzamentos através da utilização de marcadores moleculares e validar marcas específicas associadas à apomixia. Foram identificados 16 híbridos em duas populações de Brachiaria pertencentes ao programa de melhoramento. Dois marcadores associados à apomixia apresentaram amplificação conforme o esperado e foram utilizados para identificação de cinco híbridos possivelmente apomíticos. Os marcadores moleculares mostraram-se eficientes para identificação de híbridos, e poderão auxiliar a identificação precoce de materiais apomíticos, dessa forma poderão ser utilizados amplamente na seleção dentro dos programas de melhoramento