Uptake of endocytic markers into mitotic yeast cells

Abstract[3H]α-Factor and Lucifer yellow were used to measure receptor mediated and fluid-phase endocytosis in the yeast Saccharomyces cerevisiae, arrested in mitosis by depolymerization of the microtubules or due to a mutation preventing nuclear division (cdc16. Both processes continued at roughly the same level as during interphase. This shows that in yeast endocytosis is not interrupted during mitosis like in mammalian cells

Unassisted translocation of large polypeptide domains across phospholipid bilayers

Although transmembrane proteins generally require membrane-embedded machinery for integration, a few can insert spontaneously into liposomes. Previously, we established that the tail-anchored (TA) protein cytochrome b(5) (b5) can posttranslationally translocate 28 residues downstream to its transmembrane domain (TMD) across protein-free bilayers (Brambillasca, S., M. Yabal, P. Soffientini, S. Stefanovic, M. Makarow, R.S. Hegde, and N. Borgese. 2005. EMBO J. 24:2533–2542). In the present study, we investigated the limits of this unassisted translocation and report that surprisingly long (85 residues) domains of different sequence and charge placed downstream of b5's TMD can posttranslationally translocate into mammalian microsomes and liposomes at nanomolar nucleotide concentrations. Furthermore, integration of these constructs occurred in vivo in translocon-defective yeast strains. Unassisted translocation was not unique to b5 but was also observed for another TA protein (protein tyrosine phosphatase 1B) whose TMD, like the one of b5, is only moderately hydrophobic. In contrast, more hydrophobic TMDs, like synaptobrevin's, were incapable of supporting unassisted integration, possibly because of their tendency to aggregate in aqueous solution. Our data resolve long-standing discrepancies on TA protein insertion and are relevant to membrane evolution, biogenesis, and physiology

Glycosylation of rat NGF receptor ectodomain in the yeast Saccharomyces cerevisiae

AbstractHere we studied the glycosylation of a mammalian protein, the ectodomain of rat nerve growth factor receptor (NGFRe), in Saccharomyces cerevisiae. NGFRe is secreted to the culture medium of S. cerevisiae if it is fused to a polypeptide (hsp150Δ) carrier. The hsp150Δ-carrier has 95 serine and threonine residues, which were extensively O-glycosylated. In spite of 41 potential sites, NGFe lacked O-glycans, whether fused to the carrier or not. Distortion of the conformation of NGFRe by inhibition of disulfide formation did not promote O-glycosylation, whereas N-glycosylation was enhanced. Thus, the serine and threonine residues of the hsp150Δ-NGFRe fusion protein were highly selectively O-glycosylated

JRC Implementation Review 2017: In the context of the interim evaluation of the Horizon 2020 Programme

This report presents an external assessment of the follow-up that the JRC has given to the ex-post evaluation of its direct actions under the Seventh Framework Programme (FP7) conducted in 2015. The report addresses the new JRC 2030 strategy and the adaptation of the organisational structure in July 2016. The evaluation panel compliments the organisation and its leadership on the work in the design of the strategy, in gathering the support and adapting the organisational structure for the implementation of the strategy. In relatively short time the JRC went through the biggest change since the introduction of its policy-support mission in the Fifth Framework Programme in 1998. The panel notes the enthusiasm and the impressive speed with which the JRC has implemented a large number of improvements (Chapter 2). Having seen so many parts of the renewed organisation, the panel also had a critical look at the change (Chapter 3), and gives three general recommendations for further development of the JRC (Chapter 4). - Keep focus on excellence in science - Connect the whole organisation to the transformation - A modern JRC merits modern governanceJRC.DDG2-Deputy Director-General in charge of Directorates B,C,D,E,F,

EX-POST EVALUATION OF THE DIRECT ACTIONS OF THE JOINT RESEARCH CENTRE UNDER THE SEVENTH FRAMEWORK PROGRAMMES 2007-2013

The ex-post evaluation in this report provides the independent assessment requested in the Council Decisions concerning the specific programmes to be carried out by means of direct actions by the Joint Research Centre implementing the Seventh Framework Programmes (2007-2013) of the European Community and of the European Atomic Energy Community (Euratom). The evaluation has been conducted by a panel of independent external experts under the chairmanship of Professor Patrick Cunningham. In this report the Panel concludes positively on the effectiveness of the JRC as the Commission’s science service in support of Euratom and EU policies. It also concludes that the JRC has a respectable scientific performance in its areas of competence. In particular, the JRC standard is high as regards the scientific quality and impact of its publications. Besides a number of recommendations for incremental improvement of the JRC the Panel also flags two issues with a view to transformative change of the JRC. To begin with the JRC should establish a long-term strategy before the mid-term evaluation of the Horizon 2020 framework programme in 2017. As the JRC further develops its function as scientific service of the Commission, there is a need to address the JRC’s governance as well as its interaction with the scientific community in the Member States. In light of this the Commission should task a Group of eminent personalities to put forward options for JRC governance, adapted to its functions of the future. These include scientific support, research, scientific advice, and knowledge management in partnership with the Member StatesJRC.ADV02-Adviser for Evaluation and Scientific Integrit

EX-POST EVALUATION OF THE DIRECT ACTIONS OF THE JOINT RESEARCH CENTRE UNDER THE SEVENTH FRAMEWORK PROGRAMMES 2007-2013

The ex-post evaluation in this report provides the independent assessment requested in the Council Decisions concerning the specific programmes to be carried out by means of direct actions by the Joint Research Centre implementing the Seventh Framework Programmes (2007-2013) of the European Community and of the European Atomic Energy Community (Euratom). The evaluation has been conducted by a panel of independent external experts under the chairmanship of Professor Patrick Cunningham. In this report the Panel concludes positively on the effectiveness of the JRC as the Commission’s science service in support of Euratom and EU policies. It also concludes that the JRC has a respectable scientific performance in its areas of competence. In particular, the JRC standard is high as regards the scientific quality and impact of its publications. Besides a number of recommendations for incremental improvement of the JRC the Panel also flags two issues with a view to transformative change of the JRC. To begin with the JRC should establish a long-term strategy before the mid-term evaluation of the Horizon 2020 framework programme in 2017. As the JRC further develops its function as scientific service of the Commission, there is a need to address the JRC’s governance as well as its interaction with the scientific community in the Member States. In light of this the Commission should task a Group of eminent personalities to put forward options for JRC governance, adapted to its functions of the future. These include scientific support, research, scientific advice, and knowledge management in partnership with the Member StatesJRC.ADV02-Adviser for Evaluation and Scientific Integrit

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Kirjoitus perustuu esitelmään Tieteen päivillä 2005, sessiossa "Tieteiden väliset suhteet" (13.1.2005