{"id":{"repo_id":"ottawa-retro","oai_identifier":"oai:ruor.uottawa.ca:10393/19998"},"canonical_url":"https://search.dev.ndltd.org/etd/ottawa-retro/oai:ruor.uottawa.ca:10393/19998","repository":{"repo_id":"ottawa-retro","name":"University of Ottawa","base_url":"https://ruor.uottawa.ca/server/oai/request"},"display":{"title":"Understanding C/EBPbeta LAP/LIP Transcriptional and Adipogenic Potential Through Regulation by HDAC1 and GCN5","abstract":"The CCAAT/Enhancer Binding Protein Beta (C/EBPβ) is part of the leucine zipper family of transcription factors and is involved in a myriad of processes including cellular proliferation and differentiation. C/EBPβ is expressed as three isoforms (LAP*, LAP, LIP), translated from a single mRNA by a leaky ribosomal scanning mechanism. While LAP* and LAP have activating functions, LIP is recognized as being a repressor of transcription due to its lack of activation domains. Numerous studies have shown that C/EBPβ acetylation state modulates its activity in a promoter-specific manner. For instance, the acetyltransferases GCN5/PCAF and the deacetylase complex mSin3A/HDAC1 regulate C/EBPβ activity on the C/EBPa promoter. GCN5/PCAF-mediated acetylation of C/EBPβ was shown to positively affect its transcriptional activity in a steroid-dependent mechanism via the glucocorticoid receptor (GR). GR relieves HDAC1 association from C/EBPβ by targeting the deacetylase for proteasomal degradation, hence favouring GCN5-mediated acetylation of C/EBPβ and allowing maximum activation capacity to be reached. In order to further elucidate C/EBPβ activation, I sought to characterize the interplay between GCN5 and HDAC1 in regulating C/EBPβ LAP/LIP activity during murine adipogenesis by identifying their binding domain in C/EBPβ. I identified a minimal domain located within regulatory domain 1 (RD1) of C/EBPβ that is required for both GCN5 and HDAC1 binding. Furthermore, the loss of the identified domain in C/EBPβ appears to partially mimic the GR effect, thus giving C/EBPβ a higher basal transcriptional activity that accelerates NIH 3T3 and 3T3 L1 adipogenesis. Moreover, I also showed that the LIP isoform inhibitory mode of action is partially mediated through the mSin3A/HDAC1 repressor complex, which gives LIP an active repressor function. In addition to LIP inhibitory function, I also showed that a cysteine residue located in LAP* negatively regulates its transactivating function during murine adipogenesis. Although RD1 of C/EBPβ has been suggested to act as a negative regulatory domain, I showed that only five residues are responsible for most of its inhibitory effect. Hence, in an attempt to further define sub-domains within RD1, I characterized a new positive regulatory domain at its N-terminal region, which seems to be required for C/EBPβ activity in a promoter-specific manner. In conclusion, this study not only supports previously hypothesized mechanisms by which C/EBPβ is regulated, but it also redefines the contribution of LAP*, LAP and LIP in regulating transcription. Most importantly, the results emphasize the countless possibilities by which C/EBPβ transactivation potential could be modulated during cellular differentiation.","abstract_html":"The CCAAT/Enhancer Binding Protein Beta (C/EBPβ) is part of the leucine zipper family of transcription factors and is involved in a myriad of processes including cellular proliferation and differentiation. C/EBPβ is expressed as three isoforms (LAP*, LAP, LIP), translated from a single mRNA by a leaky ribosomal scanning mechanism. While LAP* and LAP have activating functions, LIP is recognized as being a repressor of transcription due to its lack of activation domains. Numerous studies have shown that C/EBPβ acetylation state modulates its activity in a promoter-specific manner. For instance, the acetyltransferases GCN5/PCAF and the deacetylase complex mSin3A/HDAC1 regulate C/EBPβ activity on the C/EBPa promoter. GCN5/PCAF-mediated acetylation of C/EBPβ was shown to positively affect its transcriptional activity in a steroid-dependent mechanism via the glucocorticoid receptor (GR). GR relieves HDAC1 association from C/EBPβ by targeting the deacetylase for proteasomal degradation, hence favouring GCN5-mediated acetylation of C/EBPβ and allowing maximum activation capacity to be reached. In order to further elucidate C/EBPβ activation, I sought to characterize the interplay between GCN5 and HDAC1 in regulating C/EBPβ LAP/LIP activity during murine adipogenesis by identifying their binding domain in C/EBPβ. I identified a minimal domain located within regulatory domain 1 (RD1) of C/EBPβ that is required for both GCN5 and HDAC1 binding. Furthermore, the loss of the identified domain in C/EBPβ appears to partially mimic the GR effect, thus giving C/EBPβ a higher basal transcriptional activity that accelerates NIH 3T3 and 3T3 L1 adipogenesis. Moreover, I also showed that the LIP isoform inhibitory mode of action is partially mediated through the mSin3A/HDAC1 repressor complex, which gives LIP an active repressor function. In addition to LIP inhibitory function, I also showed that a cysteine residue located in LAP* negatively regulates its transactivating function during murine adipogenesis. Although RD1 of C/EBPβ has been suggested to act as a negative regulatory domain, I showed that only five residues are responsible for most of its inhibitory effect. Hence, in an attempt to further define sub-domains within RD1, I characterized a new positive regulatory domain at its N-terminal region, which seems to be required for C/EBPβ activity in a promoter-specific manner. In conclusion, this study not only supports previously hypothesized mechanisms by which C/EBPβ is regulated, but it also redefines the contribution of LAP*, LAP and LIP in regulating transcription. Most importantly, the results emphasize the countless possibilities by which C/EBPβ transactivation potential could be modulated during cellular differentiation.","abstract_has_math":false,"creators":["Salem Abdou, Houssein"],"institution":"Université d'Ottawa / University of Ottawa","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":["Haché, Robert"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-17T18:17:33Z","date_published":"2011-05-17T18:17:33Z","updated_at":"2026-07-24T03:39:36Z","subjects":["adipogenesis","preadipocyte differentiation","c/ebpbeta","mSin3A/HDAC1","GCN5","transcription regulation","3T3L1 NIH3T3"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["http://dx.doi.org/10.20381/ruor-4609"],"render_values":[{"text":"http://dx.doi.org/10.20381/ruor-4609","href":"http://dx.doi.org/10.20381/ruor-4609","code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/10393/19998","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Haché, Robert"]},{"key":"dc:creator","label":"Author","values":["Salem Abdou, Houssein"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-17T18:17:33Z","2011"]},{"key":"dc:publisher","label":"Institution","values":["Université d'Ottawa / University of Ottawa"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["adipogenesis","preadipocyte differentiation","c/ebpbeta","mSin3A/HDAC1","GCN5","transcription regulation","3T3L1 NIH3T3"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/10393/19998","http://dx.doi.org/10.20381/ruor-4609"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The CCAAT/Enhancer Binding Protein Beta (C/EBPβ) is part of the leucine zipper family of transcription factors and is involved in a myriad of processes including cellular proliferation and differentiation. C/EBPβ is expressed as three isoforms (LAP*, LAP, LIP), translated from a single mRNA by a leaky ribosomal scanning mechanism. While LAP* and LAP have activating functions, LIP is recognized as being a repressor of transcription due to its lack of activation domains. Numerous studies have shown that C/EBPβ acetylation state modulates its activity in a promoter-specific manner. For instance, the acetyltransferases GCN5/PCAF and the deacetylase complex mSin3A/HDAC1 regulate C/EBPβ activity on the C/EBPa promoter. GCN5/PCAF-mediated acetylation of C/EBPβ was shown to positively affect its transcriptional activity in a steroid-dependent mechanism via the glucocorticoid receptor (GR). GR relieves HDAC1 association from C/EBPβ by targeting the deacetylase for proteasomal degradation, hence favouring GCN5-mediated acetylation of C/EBPβ and allowing maximum activation capacity to be reached. In order to further elucidate C/EBPβ activation, I sought to characterize the interplay between GCN5 and HDAC1 in regulating C/EBPβ LAP/LIP activity during murine adipogenesis by identifying their binding domain in C/EBPβ. I identified a minimal domain located within regulatory domain 1 (RD1) of C/EBPβ that is required for both GCN5 and HDAC1 binding. Furthermore, the loss of the identified domain in C/EBPβ appears to partially mimic the GR effect, thus giving C/EBPβ a higher basal transcriptional activity that accelerates NIH 3T3 and 3T3 L1 adipogenesis. Moreover, I also showed that the LIP isoform inhibitory mode of action is partially mediated through the mSin3A/HDAC1 repressor complex, which gives LIP an active repressor function. In addition to LIP inhibitory function, I also showed that a cysteine residue located in LAP* negatively regulates its transactivating function during murine adipogenesis. Although RD1 of C/EBPβ has been suggested to act as a negative regulatory domain, I showed that only five residues are responsible for most of its inhibitory effect. Hence, in an attempt to further define sub-domains within RD1, I characterized a new positive regulatory domain at its N-terminal region, which seems to be required for C/EBPβ activity in a promoter-specific manner. In conclusion, this study not only supports previously hypothesized mechanisms by which C/EBPβ is regulated, but it also redefines the contribution of LAP*, LAP and LIP in regulating transcription. Most importantly, the results emphasize the countless possibilities by which C/EBPβ transactivation potential could be modulated during cellular differentiation."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Understanding C/EBPbeta LAP/LIP Transcriptional and Adipogenic Potential Through Regulation by HDAC1 and GCN5"]}]}],"canonical_facts":{"dc:contributor":["Haché, Robert"],"dc:creator":["Salem Abdou, Houssein"],"dc:date":["2011-05-17T18:17:33Z","2011"],"dc:description":["The CCAAT/Enhancer Binding Protein Beta (C/EBPβ) is part of the leucine zipper family of transcription factors and is involved in a myriad of processes including cellular proliferation and differentiation. C/EBPβ is expressed as three isoforms (LAP*, LAP, LIP), translated from a single mRNA by a leaky ribosomal scanning mechanism. While LAP* and LAP have activating functions, LIP is recognized as being a repressor of transcription due to its lack of activation domains. Numerous studies have shown that C/EBPβ acetylation state modulates its activity in a promoter-specific manner. For instance, the acetyltransferases GCN5/PCAF and the deacetylase complex mSin3A/HDAC1 regulate C/EBPβ activity on the C/EBPa promoter. GCN5/PCAF-mediated acetylation of C/EBPβ was shown to positively affect its transcriptional activity in a steroid-dependent mechanism via the glucocorticoid receptor (GR). GR relieves HDAC1 association from C/EBPβ by targeting the deacetylase for proteasomal degradation, hence favouring GCN5-mediated acetylation of C/EBPβ and allowing maximum activation capacity to be reached. In order to further elucidate C/EBPβ activation, I sought to characterize the interplay between GCN5 and HDAC1 in regulating C/EBPβ LAP/LIP activity during murine adipogenesis by identifying their binding domain in C/EBPβ. I identified a minimal domain located within regulatory domain 1 (RD1) of C/EBPβ that is required for both GCN5 and HDAC1 binding. Furthermore, the loss of the identified domain in C/EBPβ appears to partially mimic the GR effect, thus giving C/EBPβ a higher basal transcriptional activity that accelerates NIH 3T3 and 3T3 L1 adipogenesis. Moreover, I also showed that the LIP isoform inhibitory mode of action is partially mediated through the mSin3A/HDAC1 repressor complex, which gives LIP an active repressor function. In addition to LIP inhibitory function, I also showed that a cysteine residue located in LAP* negatively regulates its transactivating function during murine adipogenesis. Although RD1 of C/EBPβ has been suggested to act as a negative regulatory domain, I showed that only five residues are responsible for most of its inhibitory effect. Hence, in an attempt to further define sub-domains within RD1, I characterized a new positive regulatory domain at its N-terminal region, which seems to be required for C/EBPβ activity in a promoter-specific manner. In conclusion, this study not only supports previously hypothesized mechanisms by which C/EBPβ is regulated, but it also redefines the contribution of LAP*, LAP and LIP in regulating transcription. Most importantly, the results emphasize the countless possibilities by which C/EBPβ transactivation potential could be modulated during cellular differentiation."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/10393/19998","http://dx.doi.org/10.20381/ruor-4609"],"dc:language":["en"],"dc:publisher":["Université d'Ottawa / University of Ottawa"],"dc:subject":["adipogenesis","preadipocyte differentiation","c/ebpbeta","mSin3A/HDAC1","GCN5","transcription regulation","3T3L1 NIH3T3"],"dc:title":["Understanding C/EBPbeta LAP/LIP Transcriptional and Adipogenic Potential Through Regulation by HDAC1 and GCN5"],"dc:type":["Thesis"]},"updated_at":"2026-07-24T03:39:36Z"}