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Download Expression of yolk protein genes in liver Beekman, Johanna
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University of Groningen Expression of yolk protein genes in liver Beekman, Johanna Margaretha IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 1990 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Beekman, J. M. (1990). Expression of yolk protein genes in liver s.n. Copyright Other than for strictly personal use, it is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum. Download date: 03-08-2017 93 Summary Themainsubject of this thesis is the transcriptionalregulationof tho chickenvitellogeninll (Vtg) and apo very low density lipoproteinll (apoVLDLll) genes. These genes code for (precursor) yolk proteins and are only expressedin the liver of a layinghen. The transcription is dependenton tho fomalesex hormoneoestradiol.In the completely oviductthe synthesis of egg white proteins is also controlledby oestradiol.The obsorvationthat two different sets of genes are under controlof the samehormonebut expressedin different tissues gives rise to one of the mainquestion ]n biology;what is the mechanism of tissue-specificand hormone-induced expressionof eukaryotic gones?. Chapter1 briefly reviewsthe regulationof gene expressionin eukaryotes.The other chaptersdescriberesults obtainedin experiments to unravelthe mechanism of oestrogen-inducedand liver-specific genes in chicken. expression of the Vtg and apoVLDLll ln Chapter2 we describethe isolationand cultivationof chicken hepatocytes(CEHcells). Thesecells transcribe their embryonic Vtg gene in an oestradiol-dependentway. By a simple endogenous calcium-phosphate coprecipitationmethodDNAcan be introducedinto thosecells and expressed.lncreasingthe oestrogen receptor concentrationartificiatly,makesthese cells a good host for gene transfer experimentsin the study of oestrogen-regulatedgene expression.Weused the CEHcells to identify regulatory regions of the Vtg gene (Chapter3). In this chapter we confirmthat the oestrogen responseelementaround position -620 is involvodin the oestrogeninducibleexpressionof the Vtg gene. This is not the only oestrogen responsiveregion identified.Another region harboursoestrogeninducibility,althoughan EREsequencecannot be discoveredin this region.Sincethis region doos not function in another oestrogen responsivecell, we do not believethat the oestrogen receptor can bind to this rogion directly. lt either needs the help of another protein to bindin this region or the other protein functions as a "bridge"to bring the receptor that binds to the EREaround position -620 close to the promoter.ln this chapter we also describethe presencoof an enhancer regionthat may bind a NFl-like protein and/or a C/EBP-likeprotein. Chapters4 and 5 describe the gene transfer experimentsperformedin the search for regulatory elementsof the apoVLDLllgen. Analysisof the 94 S'-Ílanking region revealed the presonce of two oestrogen response elom€nts,that are necessary for the oestrogen-inducible expression of the coupled CAT gene. Several other sequences appear to be involved. Described are, el€mentsthat may bind the liver-specific transcription Íactors HNF-1 or LF-A1, or elementsthat bind the CCAAT/enhancer binding protein (Chapter 6). It is clear from tho rosults presented in this thesis that regulation of gene expression is a complexprocess, that involves transcription factors that can bind to regulatory regions in several genes. The specific combinationof binding sites and the presence of specific transcription factors in the cell probably determine the strict tissuespocific, developmental-stagespecific or inducible expression of a gene. The study on the mechanismsof gene regulation has made an enormous progress for the last decade. Increasingour knowledgeon gene regulation at the molecularlevel is not only intellectually satisfying, but also vitally important in the battle against diseases such as cancer, in which tho control of gene expression is altered or lost.