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- Hsp70 and Hsp110 chaperones promote early steps of proteasome assemblyPublication . A. C., Matias; Matos, Joao; Dohmen, R. Jürgen; Ramos, Paula C.Whereas assembly of the 20S proteasome core particle (CP) in prokaryotes apparently occurs spontaneously, the efficiency of this process in eukaryotes relies on the dedicated assembly chaperones Ump1, Pba1-Pba2, and Pba3-Pba4. For mammals, it was reported that CP assembly initiates with formation of a complete alpha-ring that functions as a template for beta subunit incorporation. By contrast, we were not able to detect a ring composed only of a complete set of alpha subunits in S. cerevisiae. Instead, we found that the CP subunits alpha 1, alpha 2, and alpha 4 each form independent small complexes. Purification of such complexes containing alpha 4 revealed the presence of chaperones of the Hsp70/Ssa and Hsp110/Sse families. Consistently, certain small complexes containing alpha 1, alpha 2, and alpha 4 were not formed in strains lacking these chaperones. Deletion of the SSE1 gene in combination with deletions of PRE9 (alpha 3), PBA3, or UMP1 genes resulted in severe synthetic growth defects, high levels of ubiquitin-conjugates, and an accumulation of distinct small complexes with alpha subunits. Our study shows that Hsp70 and Hsp110 chaperones cooperate to promote the folding of individual alpha subunits and/or their assembly with other CP subunits, Ump1, and Pba1-Pba4 in subsequent steps.
- Engaging children in geosciences through storytelling and creative dancePublication . Matias, Ana Catarina; Carrasco, A. Rita; Ramos, Ana A.; Borges, R.Natural sciences have traditionally been disseminated in outreach activities as formal, one-way presentations. Nevertheless, innovative strategies are being increasingly developed using arts, gaming, and sketching, amongst others. This work aimed at testing an alternative and innovative way to engage non-expert audiences in ocean and coastal geology through a combination of scientific concept explanations and creative dancing. An informal education activity focusing on ocean dynamics was designed for 10-yearold students. It combines coastal science concepts (wind, waves, currents, and sand), storytelling techniques (narrative arc), and creative dance techniques (movement, imaginative play, and sensory engagement). A sequence of six exercises was proposed, starting with the generation of offshore ocean waves and ending with sediment transport on the beach during storm/fair-weather conditions. Scientific concepts were then translated into structured creative movements, within imaginary scenarios, and accompanied by sounds or music. The activity was performed six times with a total of 112 students. It was an inclusive activity given that all students in the class participated, including children with several mild types of cognitive and neurological impairment. The science and art activity aroused emotions of enjoyment and pleasure and allowed for effective communication between scientists and school community. Moreover, the results provide evidence of the activity’s effectiveness in engaging children and developing their willingness to further participate in similar activities.
- Prediction of overwash in alongshore variable barrier islands [Previsão da ocorrência de galgamentos em ilhas barreira com variabilidade longilitoral]Publication . Matias, Ana Catarina; Carrasco, A. Rita; Loureiro, Carlos; Almeida, S.; Ferreira, OscarOverwash prediction is very important for coastal zone management. This work intends to identify alongshore variations in storm impact and evaluate the role of sub-aerial and submerged morphologies in overwash occurrence. For this study, 24 cross-shore topo-bathymetric profiles were set on Barreta Island (Ria Formosa barrier island system, Portugal). Pre- and post-overwash surveys were made between August 2012 and April 2013. During overwash events, tidal levels and wave parameters at breaking were obtained. Overwash occurred under storm and non-storm conditions, the latter coincident with spring high-tide. Beach morphology was spatially variable, and changeable from one overwash episode to the next. Predictions of overwash occurrence were made using the Overwash Potential, defined as the difference between runup and barrier elevation. Several runup equations were tested, and the results compared to the actual observations. The selected predictor provided an accuracy of 88% for the identification of the locations where overwash occurred. This study proves that nearshore and foreshore morphologies have a major impact on the longshore distribution of overwash.
- CITED2 cooperates with ISL1 and promotes cardiac differentiation of mouse embryonic stem cellsPublication . Pacheco-Leyva, Ivette; Matias, Ana Catarina; Oliveira, Daniel V.; Santos, João; Nascimento, Rita; Guerreiro, Eduarda; Michell, Anna C.; van De Vrugt, Annebel M.; Machado-Oliveira, Gisela; Ferreira, Guilherme; Domian, Ibrahim; Bragança, JoséThe transcriptional regulator CITED2 is essential for heart development. Here, we investigated the role of CITED2 in the specification of cardiac cell fate from mouse embryonic stem cells (ESC). The overexpression of CITED2 in undifferentiated ESC was sufficient to promote cardiac cell emergence upon differentiation. Conversely, the depletion of Cited2 at the onset of differentiation resulted in a decline of ESC ability to generate cardiac cells. Moreover, loss of Cited2 expression impairs the expression of early mesoderm markers and cardiogenic transcription factors (Isl1, Gata4, Tbx5). The cardiogenic defects in Cited2-depleted cells were rescued by treatment with recombinant CITED2 protein. We showed that Cited2 expression is enriched in cardiac progenitors either derived from ESC or mouse embryonic hearts. Finally, we demonstrated that CITED2 and ISL1 proteins interact physically and cooperate to promote ESC differentiation toward cardiomyocytes. Collectively, our results show that Cited2 plays a pivotal role in cardiac commitment of ESC.
- Ectopic expression of CITED2 prior to reprogramming, promotes and homogenises the conversion of somatic cells into induced pluripotent stem cellsPublication . Charneca, João; Matias, Ana Catarina; Escapa, Ana Luisa; Fernandes, Catarina; Alves, Andre; Santos, João; Nascimento, Rita; Bragança, JoséCited2 plays crucial roles in mouse embryonic stem cells self-renewal, the initiation of the somatic reprogramming process into induced pluripotent stem cells (iPSC) and the suppression of cell senescence. Here, we investigated the potential of CITED2 expression in combination with the Oct4, Sox2, Klf4 and c-Myc factors for reprogramming of primary mouse embryonic fibroblasts (MEF) at passage 2 and 4. The ectopic CITED2 expression in primary MEF prior to the onset of the reprogramming process, generated iPSC with less variability in the expression of endogenous pluripotency-related genes. In contrast, part of the MEF reprogrammed without ectopic expression of CITED2 at passage 4 originated partially reprogrammed iPSC or pre-iPSC. However, the overexpression of CITED2 in the pre-iPSC was insufficient to complete the reprogramming process into iPSC. These results indicated that ectopic CITED2 expression at the onset of the reprogramming process in combination with the reprogramming factors promotes a complete and homogeneous conversion of somatic cells into iPSC.
- Acute Loss of Cited2 Impairs Nanog Expression and Decreases Self-Renewal of Mouse Embryonic Stem CellsPublication . Kranc, Kamil R.; Oliveira, Daniel; Armesilla-Diaz, Alejandro; Pacheco-Leyva, Ivette; Matias, Ana Catarina; Escapa, Ana Luísa; Subramani, Chithra; Wheadon, Helen; Trindade, Marlene; Nichols, Jennifer; Kaji, Keisuke; Enver, Tariq; Bragança, JoséIdentifying novel players of the pluripotency gene regulatory network centered on Oct4, Sox2, and Nanog as well as delineating the interactions within the complex network is key to understanding self-renewal and early cell fate commitment of embryonic stem cells (ESC). While overexpression of the transcriptional regulator Cited2 sustains ESC pluripotency, its role in ESC functions remains unclear. Here, we show that Cited2 is important for proliferation, survival, and self-renewal of mouse ESC. We position Cited2 within the pluripotency gene regulatory network by defining Nanog, Tbx3, and Klf4 as its direct targets. We also demonstrate that the defects caused by Cited2 depletion are, at least in part, rescued by Nanog constitutive expression. Finally, we demonstrate that Cited2 is required for and enhances reprogramming of mouse embryonic fibroblasts to induced pluripotent stem cells.