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Identifier uch.biology.msc//2004kagiabakis
Title In vivo ανάλυση πρωτεϊνικών αλληλεπιδράσεων του χαλκορυθμιζόμενου μεταγραφικού παράγοντα Mac1p της ζύμης
Author Καγιαμπάκης, Ιωάννης
Abstract This study focuses mainly on the interaction between proteins Dog2p, Yol022cp, and Rad9p with the transcription factor Mac1p. In addition, the role of those interactions was investigated. We showed that Dog2p and Rad9p interact with Mac1p in vivo by coimmunoprecipitation assays. However, using these assays it was not clear whether the Yol022cp interacted with Mac1p, thus another approach is necessary to establish a potential interaction. SDS-PAGE and monoclonal anti-phosphoserine antibody methods were used to investigate whether Dog2p affects the phosphorylation state of Mac1p, and probe the conditions of degradation Mac1p-9Myc. The latter protein did not appear to degrade in a high-copper environment, contrary to previous reports. We were unable to establish the dependence of the phosphorylation state of Mac1p on Dog2p, because the discretion of the two forms of Mac1p was very low in an SDS-PAGE. The deletion of Dog1p and Dog2p did not significantly affect transcription of CTR1 at high or low copper environments in YPD. In addition deletion of these genes did not affect intracellular concentrations of H2O2 and O2-, as did deletion of rad9. Subcellular localization of Dog2p was investigated by immunoflorescence and confocal microscopy. Dog2p was present throughout the cell, in agreement with previous record. Finally, subcloning of DOG2 in the pYX142 vector, a construct necessary for Chromatin Immunoprecipitation (CHIP) experiments, was described.
Language Greek
Issue date 2004-12-22
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
Permanent Link https://elocus.lib.uoc.gr//dlib/1/f/d/metadata-dlib-2004kagiabakis.tkl Bookmark and Share
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