BRCA1/BARD1 ubiquitinates PCNA in unperturbed conditions to promote continuous DNA synthesis.

Deficiencies in the BRCA1 tumor suppressor gene are the main cause of hereditary breast and ovarian cancer. BRCA1 is involved in the Homologous Recombination DNA repair pathway and, together with BARD1, forms a heterodimer with ubiquitin E3 activity. The relevance of the BRCA1/BARD1 ubiquitin E3 activity for tumor suppression and DNA repair remains controversial. Here, we observe that the BRCA1/BARD1 ubiquitin E3 activity is not required for Homologous Recombination or resistance to Olaparib. Using TULIP2 methodology, which enables the direct identification of E3-specific ubiquitination substrates, we identify substrates for BRCA1/BARD1. We find that PCNA is ubiquitinated by BRCA1/BARD1 in unperturbed conditions independently of RAD18. PCNA ubiquitination by BRCA1/BARD1 avoids the formation of ssDNA gaps during DNA replication and promotes continuous DNA synthesis. These results provide additional insight about the importance of BRCA1/BARD1 E3 activity in Homologous Recombination.

SUMO-activated target traps (SATTs) enable the identification of a comprehensive E3-specific SUMO proteome.

Ubiquitin and ubiquitin-like conjugation cascades consist of dedicated E1, E2, and E3 enzymes with E3s providing substrate specificity. Mass spectrometry-based approaches have enabled the identification of more than 6500 SUMO2/3 target proteins. The limited number of SUMO E3s provides the unique opportunity to systematically study E3 substrate wiring. We developed SUMO-activated target traps (SATTs) and systematically identified substrates for eight different SUMO E3s, PIAS1, PIAS2, PIAS3, PIAS4, NSMCE2, ZNF451, LAZSUL (ZNF451-3), and ZMIZ2. SATTs enabled us to identify 427 SUMO1 and 961 SUMO2/3 targets in an E3-specific manner. We found pronounced E3 substrate preference. Quantitative proteomics enabled us to measure substrate specificity of E3s, quantified using the SATT index. Furthermore, we developed the Polar SATTs web-based tool to browse the dataset in an interactive manner. Overall, we uncover E3-to-target wiring of 1388 SUMO substrates, highlighting unique and overlapping sets of substrates for eight different SUMO E3 ligases.

Ubiquitinome Profiling Reveals in Vivo UBE2D3 Targets and Implicates UBE2D3 in Protein Quality Control.

Ubiquitination has crucial roles in many cellular processes, and dysregulation of ubiquitin machinery enzymes can result in various forms of pathogenesis. Cells only have a limited set of ubiquitin-conjugating (E2) enzymes to support the ubiquitination of many cellular targets. As individual E2 enzymes have many different substrates and interactions between E2 enzymes and their substrates can be transient, it is challenging to define all in vivo substrates of an individual E2 and the cellular processes it affects. Particularly challenging in this respect is UBE2D3, an E2 enzyme with promiscuous activity in vitro but less defined roles in vivo. Here, we set out to identify in vivo targets of UBE2D3 by using stable isotope labeling by amino acids in cell culture-based and label-free quantitative ubiquitin diGly proteomics to study global proteome and ubiquitinome changes associated with UBE2D3 depletion. UBE2D3 depletion changed the global proteome, with the levels of proteins from metabolic pathways, in particular retinol metabolism, being the most affected. However, the impact of UBE2D3 depletion on the ubiquitinome was much more prominent. Interestingly, molecular pathways related to mRNA translation were the most affected. Indeed, we find that ubiquitination of the ribosomal proteins RPS10 and RPS20, critical for ribosome-associated protein quality control, is dependent on UBE2D3. We show by Targets of Ubiquitin Ligases Identified by Proteomics 2 methodology that RPS10 and RPS20 are direct targets of UBE2D3 and demonstrate that the catalytic activity of UBE2D3 is required to ubiquitinate RPS10 in vivo. In addition, our data suggest that UBE2D3 acts at multiple levels in autophagic protein quality control. Collectively, our findings show that depletion of an E2 enzyme in combination with quantitative diGly-based ubiquitinome profiling is a powerful tool to identify new in vivo E2 substrates, as we have done here for UBE2D3. Our work provides an important resource for further studies on the in vivo functions of UBE2D3.

Insights in Post-Translational Modifications: Ubiquitin and SUMO.

Both ubiquitination and SUMOylation are dynamic post-translational modifications that regulate thousands of target proteins to control virtually every cellular process. Unfortunately, the detailed mechanisms of how all these cellular processes are regulated by both modifications remain unclear. Target proteins can be modified by one or several moieties, giving rise to polymers of different morphology. The conjugation cascades of both modifications comprise a few activating and conjugating enzymes but close to thousands of ligating enzymes (E3s) in the case of ubiquitination. As a result, these E3s give substrate specificity and can form polymers on a target protein. Polymers can be quickly modified forming branches or cleaving chains leading the target protein to its cellular fate. The recent development of mass spectrometry(MS) -based approaches has increased the understanding of ubiquitination and SUMOylation by finding essential modified targets in particular signaling pathways. Here, we perform a concise overview comprising from the basic mechanisms of both ubiquitination and SUMOylation to recent MS-based approaches aimed to find specific targets for particular E3 enzymes.

Community Intervention Self-Efficacy Scale for Parent Leaders (CONNECTED): Parents' Empowerment to Prevent Adolescent Alcohol Use.

Empowering parents by actively engaging them in environmental prevention strategies is a promising approach that only a few programs use. Evidence suggests that when families and the wider community are engaged, alcohol prevention is more efficient. However, due to the novelty of this approach, no specific assessment tools for measuring this type of engagement are available. The objective of this study is to design a parental empowerment measurement tool to evaluate parents' self-efficacy when engaging in environmental and community actions and to analyze its psychometric properties. A total of 132 parents active in in-school parent associations from Spain (n = 77; 58.4%) and Portugal (n = 55; 41.7%) completed a pencil and paper battery of four questionnaires, including the developed scale COmmuNity iNtervention SElf-Efficacy SCale for ParenT LEaDers (CONNECTED). The scale showed a good reliability and good test-retest stability in a three-month period. The convergent validity with other well-established instruments that assess similar constructs was significant. A preliminary confirmatory factor analysis (CFA) showed an acceptable fit. Environmental prevention supported by families is a promising preventive strategy because the participation and involvement of families is an effective way to address some risks in adolescence; however, new assessment tools are needed in this field. The developed scale could be a first step to identify the areas of need in a community and to monitor the progress and evaluate the outcomes of the preventive interventions implemented.

TULIP2: An Improved Method for the Identification of Ubiquitin E3-Specific Targets.

Protein modification by Ubiquitin or Ubiquitin-like modifiers is mediated by an enzyme cascade composed of E1, E2, and E3 enzymes. E1s, or ubiquitin-activating enzymes, perform ubiquitin activation. Next, ubiquitin is transferred to ubiquitin-conjugating enzymes or E2s. Finally, ubiquitin ligases or E3s catalyze the transfer of ubiquitin to the acceptor proteins. E3 enzymes are responsible for determining the substrate specificity. Determining which E3 enzyme maps to which substrate is a major challenge that is greatly facilitated by the TULIP2 methodology. TULIP2 methodology is fast, precise, and cost-effective. Compared to the previous TULIP methodology protocol, TULIP2 methodology achieves a more than 50-fold improvement in the purification yield and two orders of magnitude improvement in the signal-to-background ratio after label free quantification by mass spectrometry analysis. The method includes the generation of TULIP2 cell lines, subsequent purification of TULIP2 conjugates, preparation, and analysis of samples by mass spectrometry.

La percepción del apoyo social en jugadores patológicos y su relación con el éxito del tratamiento / Perceived social support in pathological gamblers and treatment effectiveness

El juego patológico afecta entre el 0.4% y el 1.5% de la población mundial. Aunque existen procedimientos terapéuticos eficaces, la escasa adherencia terapéutica y las recaídas amenazan los resultados. El objetivo es determinar la relación entre el apoyo social, la adherencia terapéutica y el cese del juego. Se evaluó convivencia, acompañamiento, implicación e intervención del acompañante en una muestra de 68 jugadores patológicos. Los resultados muestran una asociación no significativa entre apoyo social y adherencia al tratamiento y éxito terapéutico. Se mantuvieron en tratamiento el 76.0% de los que tenían apoyo social alto frente al 42.1% que tenían apoyo social bajo. El éxito terapéutico presenta una asociación significativa con el estado civil. Las personas que mantienen una relación de pareja estable tienen el doble de probabilidad de finalizar el tratamiento con éxito. Los resultados sugieren considerar el apoyo social como una variable decisiva en el tratamiento de la ludopatía

Pathological gambling affects 0.4-1.5 percent of the global population. Therapy has proved effective, even though low therapeutic compliance and frequent withdrawals weakens good treatment results. The aim of this paper is to evaluate the relationship between social support, therapeutic adherence, and gambling cessation. Four aspects of social support were measured: cohabitation, patient's companion, commitment, and intervention with patient's companion, in a sample of 68 pathological gamblers. Results show a non-significant trend for social support and treatment compliance to be associated to treatment success. Seventy-six percent of high social support patients kept the treatment up to the end vis-à-vis42.1% of low social support patients. Therapeutic success shows a significant relationship to a stable couple relationship. Individuals who had stable couple relationship were twice likely to finish treatment successfully. Results suggest that social support should be considered as a critical variable in pathological gambling treatment

Meta-analysis of the effectiveness of school substance abuse prevention programs in Spain / Meta-análisis de la eficacia de los programas de prevención escolares del abuso de sustancias en España

BACKGROUND: There has been an increase in adolescent substance use that has led to the development and implementation of prevention programs. New evidence is needed in order to improve them and optimize the resources. The aim of this paper is to use a meta-analysis to analyze the effectiveness of school drug prevention programs in Spain. METHOD: Twenty-one studies that evaluated drug abuse prevention programs in schools, were published between 2002 and 2013, and that met the selection criteria were identified. RESULTS: Preventive program effectiveness was low (d= 0.16), although it was higher at the follow-up (d= 0.30). The programs were most effective in changing attitudes (d= 0.44) towards drugs. The models of health education (d= 0.48) and social learning (d= 0.20) were also very effective, especially in combination with oral, written, and audiovisual support material (d= 0.21) and the implementation of joint programs by health education professionals and faculty members (d= 0.25). CONCLUSIONS: Is possible to determine the need for more rigorous evaluations of interventions to establish useful programs

ANTECEDENTES: en los últimos años se ha producido un aumento del consumo de sustancias en población adolescente. Es necesario contar con nuevas evidencias sobre programas que funcionan y mejorar aquellos que se están realizando con el fin de optimizar los recursos. El objetivo de este trabajo es analizar la eficacia de los programas escolares de prevención de drogas en España mediante un meta-análisis. MÉTODO: se identificaron 21 estudios publicados entre 2002 y 2013 que cumplieron los criterios de selección y que evaluaban programas de prevención del abuso de drogas en el ámbito escolar en España. RESULTADOS: la eficacia preventiva de los programas fue baja (d= 0.16), aunque era mayor en las medidas de seguimiento (d= 0.30). Los programas resultaron más eficaces en la modificación de las actitudes hacia las drogas (d= 0.44). Los modelos de educación para la salud (d= 0.48) y aprendizaje social (d= 0.20) resultaron más eficaces, junto con la combinación de soporte oral, escrito y audiovisual del material (d= 0.21) y la implementación de los programas por profesionales y profesorado conjuntamente (d= 0.25). CONCLUSIONES: A partir de los resultados es posible determinar la necesidad de evaluaciones más rigorosas de las intervenciones

Meta-analysis of the effectiveness of school substance abuse prevention programs in Spain.

BACKGROUND: There has been an increase in adolescent substance use that has led to the development and implementation of prevention programs. New evidence is needed in order to improve them and optimize the resources. The aim of this paper is to use a meta-analysis to analyze the effectiveness of school drug prevention programs in Spain. METHOD: Twenty-one studies that evaluated drug abuse prevention programs in schools, were published between 2002 and 2013, and that met the selection criteria were identified. RESULTS: Preventive program effectiveness was low ( d = 0.16), although it was higher at the follow-up ( d = 0.30). The programs were most effective in changing attitudes ( d = 0.44) towards drugs. The models of health education ( d = 0.48) and social learning ( d = 0.20) were also very effective, especially in combination with oral, written, and audiovisual support material ( d = 0.21) and the implementation of joint programs by health education professionals and faculty members ( d = 0.25). CONCLUSIONS: Is possible to determine the need for more rigorous evaluations of interventions to establish useful programs.

Waking self-hypnosis efficacy in cognitive-behavioral treatment for pathological gambling: an effectiveness clinical assay.

Cognitive-behavioral therapy for pathological gambling has a long-term success rate of more than 50%. This study evaluated the effect of self-hypnosis in cognitive-behavioral treatment of pathological gamblers. Forty-nine participants were assigned to 2 groups. Both groups received a cognitive-behavioral protocol, and Group 1, the no-hypnosis group, received an 11-session intervention and Group 2, the hypnosis group, received 7 sessions that included self-hypnosis. Both groups were equal in gambling chronicity, frequency, intensity, change motivation, and problems derived from gambling. All participants reported significant improvement in gambling behavior and consequences at both treatment end and 6-month follow-up. Data show no differences between the interventions in abstinence, therapeutic compliance, fulfillment, and satisfaction. Results suggest that self-hypnosis reinforces treatment and can be a supportive technique for future brief interventions.

Applying drug dependence research to prevention interventions in Spain.

This article examines the status of evidence-based interventions for preventing drug dependence in Spain. The evolution of the perception that the Spanish have of the problem and how this has influenced prevention efforts is described. An analysis is made of how programs designed to prevent drug use have been translated from the field of experimental research to implementation. The characteristics of evidence-based programs developed in Spain are outlined, analyzing their efficacy and the adaptations of programs from other countries to the Spanish context. Most have been school based, although some family and leisure-time based programs also have been developed. The processes for translation and cultural adaptation of evidence-based programs are described. Finally, pending aspects of the adaptation of research in drug dependence within Spain are discussed.