Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor
CREB is an important transcription factor that is regulated in part by the CRTC family of cofactors. CRTCs work in tandem with CREB to mediate various metabolic processes, such as promoting gluconeogenesis in response to increased glucagon levels. CRTC1 in particular is highly expressed in the brai...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2015
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| Online Access: | http://hdl.handle.net/10356/65366 |
| _version_ | 1826112537526009856 |
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| author | Nai, Rui Si |
| author2 | Mark Stephen Featherstone |
| author_facet | Mark Stephen Featherstone Nai, Rui Si |
| author_sort | Nai, Rui Si |
| collection | NTU |
| description | CREB is an important transcription factor that is regulated in part by the CRTC family of cofactors. CRTCs work in tandem with CREB to mediate various metabolic processes, such as promoting gluconeogenesis in response to increased glucagon levels. CRTC1 in particular is highly expressed in the brain and known to be involved in memory formation, learning and synaptic plasticity. Consequently, its dysregulation is linked to neurological diseases as well as memory and learning problems. Identifying and studying novel CRTC1 partners may provide a better understanding of how CRTC1 functions. This project tests the feasibility of BioID in pulling down CRTC1 partner proteins. BioID is a protein screening technique that involves fusing a mutant E.coli biotin ligase, BirA*, to the protein of interest in order to biotinylate proximal proteins. These potential interaction partners can then be identified through streptavidin pull-down and Western blot or mass spectrometry. Results showed that known CRTC1 partners CREB and MEIS1A were successfully pulled down when overexpressed, demonstrating that BioID is functional and that novel protein partners of CRTC1 can potentially be pulled down with this method. |
| first_indexed | 2024-10-01T03:08:40Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/65366 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:08:40Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | ntu-10356/653662023-02-28T18:06:13Z Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor Nai, Rui Si Mark Stephen Featherstone School of Biological Sciences DRNTU::Science::Biological sciences::Genetics CREB is an important transcription factor that is regulated in part by the CRTC family of cofactors. CRTCs work in tandem with CREB to mediate various metabolic processes, such as promoting gluconeogenesis in response to increased glucagon levels. CRTC1 in particular is highly expressed in the brain and known to be involved in memory formation, learning and synaptic plasticity. Consequently, its dysregulation is linked to neurological diseases as well as memory and learning problems. Identifying and studying novel CRTC1 partners may provide a better understanding of how CRTC1 functions. This project tests the feasibility of BioID in pulling down CRTC1 partner proteins. BioID is a protein screening technique that involves fusing a mutant E.coli biotin ligase, BirA*, to the protein of interest in order to biotinylate proximal proteins. These potential interaction partners can then be identified through streptavidin pull-down and Western blot or mass spectrometry. Results showed that known CRTC1 partners CREB and MEIS1A were successfully pulled down when overexpressed, demonstrating that BioID is functional and that novel protein partners of CRTC1 can potentially be pulled down with this method. Bachelor of Science in Biological Sciences 2015-08-28T00:44:29Z 2015-08-28T00:44:29Z 2015 2015 Final Year Project (FYP) http://hdl.handle.net/10356/65366 en Nanyang Technological University 31 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Genetics Nai, Rui Si Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor |
| title | Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor |
| title_full | Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor |
| title_fullStr | Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor |
| title_full_unstemmed | Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor |
| title_short | Use of a promiscuous biotin ligase to identify partners of the CRTC transcription factor |
| title_sort | use of a promiscuous biotin ligase to identify partners of the crtc transcription factor |
| topic | DRNTU::Science::Biological sciences::Genetics |
| url | http://hdl.handle.net/10356/65366 |
| work_keys_str_mv | AT nairuisi useofapromiscuousbiotinligasetoidentifypartnersofthecrtctranscriptionfactor |