Quantitative Analyses of the Yeast Oxidative Protein Folding Pathway in Vitro and in Vivo

Aims: Efficient oxidative protein folding (OPF) in the endoplasmic reticulum (ER) is a key requirement of the eukaryotic secretory pathway. In particular, protein folding linked to the formation of disulfide bonds, an activity dependent on the enzyme protein disulfide isomerase (PDI), is crucial. For the de novo formation of disulphide bonds, reduced PDI must be re-oxidised by an ER-located oxidase (ERO1). Despite some knowledge of this pathway, the kinetic parameters with which these components act and the importance of specific parameters, such as PDI reoxidation by Ero1, for the overall performance of OPF in vivo remain poorly understood. Results: We established an in vitro system using purified yeast (Saccharomyces cerevisiae) PDI (Pdi1p) and ERO1 (Ero1p) to investigate OPF. This necessitated the development of a novel reduction/oxidation processing strategy to generate homogenously oxidised recombinant yeast Ero1p. This new methodology enabled the quantitative assessment of the interaction of Pdi1p and Ero1p in vitro by measuring oxygen consumption and reoxidation of reduced RNAase A. The resulting quantitative data were then used to generate a simple model which can describe the oxidising capacity of Pdi1p and Ero1p in vitro and predict the in vivo effect of modulation of the levels of these proteins. Innovation: We describe a model that can be used to explore the OPF pathway and its control in a quantitative way. Conclusion: Our study provides new insights into how OPF works at a molecular level and provides a platform for the design of more efficient heterologous protein expression systems in yeast

GODLY PLAY SUOMESSA : Kyselytutkimus Godly play -uskontokasvatusmenetelmän käyttäjille

Hakulinen, Heidi. Godly play Suomessa – Kyselytutkimus Godly play -uskontokasvatusmenetelmän käyttäjille. Diak Etelä, Järvenpää, Kevät 2011, 54s., 1 liite. Diakonia-ammattikorkeakoulu, Sosiaalialan koulutusohjelma, Kristillisen lapsi- ja nuorisotyön suuntautumisvaihtoehto, sosionomi (AMK) + kirkon nuorisotyönohjaajan virkakelpoisuus + lastentarhanopettajan kelpoisuus. Tämän opinnäytetyön tarkoitus oli selvittää millaista Godly play -toimintaa Suomessa on ja kuinka paljon menetelmää käytetään. Vastaajat tavoitettiin Seurakuntaopiston kautta, joten kysely ei tavoittanut kaikkia Suomen Godly playn käyttäjiä. Kyselylomakkeita lähetettiin 34 käyttäjälle ja vastauksia saatiin 31. Tutkimus toteutettiin kvantitatiivisesti ja tutkimusaineisto kerättiin syksyllä 2010 sähköpostitse. Aineisto analysoitiin SPSS -ohjelmalla sekä sisällön analyysi -menetelmällä talvella 2010–2011. Tutkimuksessa selvisi, että Suomessa on vielä melko vähän Godly playn käyttäjiä. Menetelmää käytetään seurakunnan monilla eri työaloilla, mutta myös seurakunnan ulkopuolella. Eniten menetelmää käytetään seurakunnan päiväkerhossa, rippikoulussa sekä aikuisille suunnatuissa hartaushetkissä. Godly playtä käytetään vain osittain sen alkuperäisessä muodossaan. Reagointityöskentelyä käytetään yllättävän vähän siihen nähden, että sen sanotaan olevan koko menetelmän tärkein osa.Hakulinen, Heidi. Godly play in Finland – a Survey for the users of Godly play religious education method. 54p,. 1 appendix. Language: Finnish. Järvenpää, Spring 2011. Diaconia University of Applied Sciences. Degree Program in Social Services, Option in Christian Youth Work. Degree: Bachelor of Social Services. The purpose of the thesis was to find out what kind of Godly play activities there are in Finland and how often this religious education method is used. The respondents were contacted through the Church Training College, so that survey didn’t reach every Godly play user in Finland. The questionnaire was sent to altogether 34 users and responses were received 31. The research was carried out quantitatively and the data were collected in autumn 2010 via e-mail. The data were analyzed using the SPSS program and content analysis method in winter 2010–2011. The result showed that there are quite few Godly play users in Finland. The method is used most in the church day care, confirmation camps and hours of devotion for adults. In addition, it is used for example in day care centres and schools. Godly play is only partly used as it originally is supposed to be used. The session includes the response time surprisingly rarely taking into account that it is considered the most important part in the whole method. Keywords: Godly play, Montessori Method, Christian education, quantitative researc

Monoculture Breeds Poor Social Skills

Two studies from Jarosz et al. describe how [GAR+], a protein-based epigenetic determinant found mainly in wild yeast strains, can be activated by microbial cross-kingdom communication. With the aid of genetically and ecologically diverse bacteria, yeast can override an ancient regulatory mechanism of glucose repression, promoting both microbial diversity and lifespan extension

[PSI+] turns 50

The year 2015 sees the fiftieth anniversary of the publication of a research paper that underpins much of our understanding of fungal prion biology, namely “?, a cytoplasmic suppressor of super-suppressor in yeast” by Brian Cox. Here we show how our understanding of the molecular nature of the [PSI+] determinant evolved from an ‘occult’ determinant to a transmissible amyloid form of a translation termination factor. We also consider the impact studies on [PSI] have had – and continue to have - on prion research. To demonstrate this, leading investigators in the yeast prion field who have made extensive use of the [PSI+] trait in their research, provide their own commentaries on the discovery and significance of [PSI]