O Yap1 no stress causado pelo excesso de cobalto em S. cerevisiae

Mestrado em BiotecnologiaA capacidade dos organismos em alterar os seus padrões de expressão de genes em resposta a perturbações do meio ambiente é essencial para a sua viabilidade. Neste trabalho, utilizando Saccharomyces cerevisiae como organismo eucariota modelo, foi estudada a relevância do factor de transcrição da família Yap, o Yap1, na desintoxicação do excesso de cobalto. Os resultados obtidos neste trabalho demonstram que, após a incubação das células com cobalto, o factor de transcrição Yap1 é acumulado no núcleo, através de um mecanismo independente de Orp1/Gpx3. Verificámos que o cobalto tem a capacidade de ligar-se directamente ao Yap1, sugerindo que, à semelhança do que acontece com outros metais, seja este o mecanismo de retenção nuclear após o stress. Confirmámos que uma vez no núcleo, o Yap1 medeia a indução de genes envolvidos na regulação do stress oxidativo em levedura (tais como GPX2, SOD1, TRR1 e TRX2) e constatámos que a presença de cobalto aumenta os níveis de carbonilação das proteínas, sendo esse aumento mais acentuado na ausência de Yap1. Verificámos ainda que os níveis de cobalto estão levemente diminuídos no mutante yap1 ao contrário do ferro que está em concentrações mais elevadas neste mutante relativamente à estirpe selvagem, quer na presença como na ausência de cobalto. Este facto leva-nos a propor que o cobalto altere a homeostase do ferro e que em consequência, conjuntamente com a diminuição das defesas anti-oxidantes, gere stress oxidativo. Foi descrito que a glutationa (GSH) é importante na homeostase do ferro e o excesso de cobalto leva ao aumento dos níveis de ferro intracelulares. Verificámos não só que a expressão de GSH1 está diminuída aos 30 minutos de tratamento do mutante yap1 com cobalto mas também que a adição de GSH faz recuperar os fenótipos de crescimento na presença deste metal. Finalmente apresentámos um modelo de acção do factor Yap1 no stress provocado pelo cobalto.The ability of organisms to reprogram gene expression in response to changes in the environment is essential for their viability. In this work, using Saccharomyces cerevisiae as a eukaryotic model organism, we studied the relevance of the transcription factor, Yap1, in the detoxification of cobalt excess. The results here reported show that after incubation of cells with cobalt, Yap1 accumulates in the nucleus, through a mechanism independent of the peroxidase Orp1/Gpx3. We found that cobalt has the ability to directly bind to Yap1, suggesting that this is the mechanism for its nuclear retention after cobalt treatment. We confirmed that, once in the nucleus, Yap1 mediates the induction of genes involved in the regulation of oxidative stress in yeast (such as GPX2, SOD1, TRX2 and TRR1). Moreover, we found that the presence of cobalt increases the levels of protein carbonylation, being the damage more pronounced in the absence of Yap1. It was also found that cobalt levels are slightly decreased in the yap1 mutant. In contrast, the iron concentration is higher in this mutant than in the wild type, in the presence and absence of cobalt. This led us to propose that the cobalt alters iron homeostasis and iin consequence generate oxidative stress. It was reported that glutathione (GSH) is important in the homeostasis of iron. As the excess of cobalt leads to increased levels of intracellular iron, we found not only that in the yap1 mutant GSH1 expression is decreased after 30 minutes of treatment with cobalt but also, the addition of GSH can recover the growth phenotypes in the presence of this metal. . Finally we present a working model of the transcription factor Yap1 in stress caused by cobalt

The cases of cobalt, cadmium and iron

" The Genomics and Stress Laboratory, at ITQB, headed by Prof. Claudina Rodrigues- Pousada, has been focused on the identification and characterization of stress-responsive genetic programs regulating metal toxicity in the yeast Saccharomyces cerevisiae(...)

An internal promoter drives the expression of a truncated form of ccc1 capable of protecting yeast from iron toxicity

Funding Information: This work was supported by Project LISBOA-01-0145-FEDER-007660 (?Microbiologia Molecular, Estrutural e Celular?) funded by FEDER funds through COMPETE2020??Programa Operacional Competitividade e Internacionaliza??o? (POCI); ?Funda??o para a Ci?ncia e a Tecnologia? (FCT) grants EXPL/BIA-MIC/2525/2013 and programme IF (IF/00124/2015) to C.P.; grant PTDC/BIA-BQM/31317/2017 to C.V.R.; the European Union?s Horizon 2020 research and innovation programme under grant agreement No. 810856 and 857203; COST Action CA15133, supported by COST (European Cooperation in Science and Technology) and PPBI?Portuguese Platform of BioImaging (PPBI-POCI-01-0145-FEDER-022122) co-funded by national funds from OE??Or?amento de Estado? and by FEDER. C.A. (SFRH/BPD/74294/2010), S.M.C. (SFRH/BD/91077/2012), C.V.R. (SFRH/BPD/94050/2013) and A.G.-C. (SFRH/BD/118866/2016) were supported by FCT contracts or fellowships. Funding Information: Funding: This work was supported by Project LISBOA-01-0145-FEDER-007660 (“Microbiologia Molecular, ?strutural e Celular”) funded by F?D?R funds through COMP?T?2020—“Programa Operacional Competitividade e Internacionalização” (POCI); “Fundação para a Ciência e a Tecno-logia” (FCT) grants ?XPL/BIA-MIC/2525/2013 and programme IF (IF/00124/2015) to C.P.; grant PTDC/BIA-BQM/31317/2017 to C.V.R.; the ?uropean Union’s Horizon 2020 research and innovation programme under grant agreement No. 810856 and 857203; COST Action CA15133, supported by COST (European Cooperation in Science and Technology) and PPBI—Portuguese Platform of Bi-oImaging (PPBI-POCI-01-0145-FEDER-022122) co-funded by national funds from OE—“Orçamento de Estado” and by FEDER. C.A. (SFRH/BPD/74294/2010), S.M.C. (SFRH/BD/91077/2012), C.V.R. (SFRH/BPD/94050/2013) and A.G.-C. (SFRH/BD/118866/2016) were supported by FCT contracts or fellowships. Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.In yeast, iron storage and detoxification depend on the Ccc1 transporter that mediates iron accumulation in vacuoles. While deletion of the CCC1 gene renders cells unable to survive under iron overload conditions, the deletion of its previously identified regulators only partially affects survival, indicating that the mechanisms controlling iron storage and detoxification in yeast are still far from well understood. This work reveals that CCC1 is equipped with a complex transcriptional structure comprising several regulatory regions. One of these is located inside the coding sequence of the gene and drives the expression of a short transcript encoding an N-terminally truncated protein, designated as s-Ccc1. s-Ccc1, though less efficiently than Ccc1, is able to promote metal accumulation in the vacuole, protecting cells against iron toxicity. While the expression of the s-Ccc1 appears to be repressed in the normal genomic context, our current data clearly demonstrates that it is functional and has the capacity to play a role under iron overload conditions.publishersversionpublishe

MAMMALS IN PORTUGAL : A data set of terrestrial, volant, and marine mammal occurrences in P ortugal

Mammals are threatened worldwide, with 26% of all species being includedin the IUCN threatened categories. This overall pattern is primarily associatedwith habitat loss or degradation, and human persecution for terrestrial mam-mals, and pollution, open net fishing, climate change, and prey depletion formarine mammals. Mammals play a key role in maintaining ecosystems func-tionality and resilience, and therefore information on their distribution is cru-cial to delineate and support conservation actions. MAMMALS INPORTUGAL is a publicly available data set compiling unpublishedgeoreferenced occurrence records of 92 terrestrial, volant, and marine mam-mals in mainland Portugal and archipelagos of the Azores and Madeira thatincludes 105,026 data entries between 1873 and 2021 (72% of the data occur-ring in 2000 and 2021). The methods used to collect the data were: live obser-vations/captures (43%), sign surveys (35%), camera trapping (16%),bioacoustics surveys (4%) and radiotracking, and inquiries that represent lessthan 1% of the records. The data set includes 13 types of records: (1) burrowsjsoil moundsjtunnel, (2) capture, (3) colony, (4) dead animaljhairjskullsjjaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8),observation in shelters, (9) photo trappingjvideo, (10) predators dietjpelletsjpine cones/nuts, (11) scatjtrackjditch, (12) telemetry and (13) vocalizationjecholocation. The spatial uncertainty of most records ranges between 0 and100 m (76%). Rodentia (n=31,573) has the highest number of records followedby Chiroptera (n=18,857), Carnivora (n=18,594), Lagomorpha (n=17,496),Cetartiodactyla (n=11,568) and Eulipotyphla (n=7008). The data setincludes records of species classified by the IUCN as threatened(e.g.,Oryctolagus cuniculus[n=12,159],Monachus monachus[n=1,512],andLynx pardinus[n=197]). We believe that this data set may stimulate thepublication of other European countries data sets that would certainly contrib-ute to ecology and conservation-related research, and therefore assisting onthe development of more accurate and tailored conservation managementstrategies for each species. There are no copyright restrictions; please cite thisdata paper when the data are used in publications.info:eu-repo/semantics/publishedVersio

Mammals in Portugal: a data set of terrestrial, volant, and marine mammal occurrences in Portugal

Mammals are threatened worldwide, with ~26% of all species being included in the IUCN threatened categories. This overall pattern is primarily associated with habitat loss or degradation, and human persecution for terrestrial mammals, and pollution, open net fishing, climate change, and prey depletion for marine mammals. Mammals play a key role in maintaining ecosystems functionality and resilience, and therefore information on their distribution is crucial to delineate and support conservation actions. MAMMALS IN PORTUGAL is a publicly available data set compiling unpublished georeferenced occurrence records of 92 terrestrial, volant, and marine mammals in mainland Portugal and archipelagos of the Azores and Madeira that includes 105,026 data entries between 1873 and 2021 (72% of the data occurring in 2000 and 2021). The methods used to collect the data were: live observations/captures (43%), sign surveys (35%), camera trapping (16%), bioacoustics surveys (4%) and radiotracking, and inquiries that represent less than 1% of the records. The data set includes 13 types of records: (1) burrows | soil mounds | tunnel, (2) capture, (3) colony, (4) dead animal | hair | skulls | jaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8), observation in shelters, (9) photo trapping | video, (10) predators diet | pellets | pine cones/nuts, (11) scat | track | ditch, (12) telemetry and (13) vocalization | echolocation. The spatial uncertainty of most records ranges between 0 and 100 m (76%). Rodentia (n =31,573) has the highest number of records followed by Chiroptera (n = 18,857), Carnivora (n = 18,594), Lagomorpha (n = 17,496), Cetartiodactyla (n = 11,568) and Eulipotyphla (n = 7008). The data set includes records of species classified by the IUCN as threatened (e.g., Oryctolagus cuniculus [n = 12,159], Monachus monachus [n = 1,512], and Lynx pardinus [n = 197]). We believe that this data set may stimulate the publication of other European countries data sets that would certainly contribute to ecology and conservation-related research, and therefore assisting on the development of more accurate and tailored conservation management strategies for each species. There are no copyright restrictions; please cite this data paper when the data are used in publications