Nuclear translation
<p>In bacteria, protein synthesis can occur tightly coupled to transcription. In eukaryotes, it is believed that translation occurs solely in the cytoplasm; I test whether some occurs in nuclei and find: (1) L-azidohomoalanine (Aha) – a methionine analogue (detected by microscopy after attachi...
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| Format: | Thesis |
| Language: | English |
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2012
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| _version_ | 1826316685786742784 |
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| author | Baboo, S |
| author2 | Cook, P |
| author_facet | Cook, P Baboo, S |
| author_sort | Baboo, S |
| collection | OXFORD |
| description | <p>In bacteria, protein synthesis can occur tightly coupled to transcription. In eukaryotes, it is believed that translation occurs solely in the cytoplasm; I test whether some occurs in nuclei and find: (1) L-azidohomoalanine (Aha) – a methionine analogue (detected by microscopy after attaching a fluorescent tag using ‘click’ chemistry) – is incorporated within 5 s into nuclei in a process sensitive to the translation inhibitor, anisomycin. (2) Puromycin – another inhibitor that end-labels nascent peptides (detected by immuno-fluorescence) – is similarly incorporated in a manner sensitive to a transcriptional inhibitor. (3) CD2 – a non-nuclear protein – is found in nuclei close to the nascent RNA that encodes it (detected by combining indirect immuno-labelling with RNA fluorescence in situ hybridization using intronic probes); faulty (nascent) RNA is destroyed by a quality-control mechanism sensitive to translational inhibitors. I conclude that substantial translation occurs in the nucleus, with some being closely coupled to transcription and the associated proof-reading. Moreover, most peptides made in both the nucleus and cytoplasm are degraded soon after they are made with half-lives of about one minute.</p>
<p>I also collaborated on two additional projects: the purification of mega-complexes (transcription 'factories’) containing RNA polymerases I, II, or III (I used immuno-fluorescence to confirm that each contained the expected constituents), and the demonstration that some ‘factories’ specialize in transcribing genes responding to tumour necrosis factor α – a cytokine that signals through NFκB (I used RNA fluorescence in situ hybridization coupled with immuno-labelling to show active NFκB is found in factories transcribing responsive genes).</p> |
| first_indexed | 2024-03-07T07:42:21Z |
| format | Thesis |
| id | oxford-uuid:5266f049-d576-44fd-ab26-11cf7a27f678 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-12-09T03:49:23Z |
| publishDate | 2012 |
| record_format | dspace |
| spelling | oxford-uuid:5266f049-d576-44fd-ab26-11cf7a27f6782024-12-08T12:38:58ZNuclear translationThesishttp://purl.org/coar/resource_type/c_db06uuid:5266f049-d576-44fd-ab26-11cf7a27f678Synthetic organic chemistryProtein chemistryCell BiologyMicroscopyBiologyEnglishOxford University Research Archive - Valet2012Baboo, SCook, P<p>In bacteria, protein synthesis can occur tightly coupled to transcription. In eukaryotes, it is believed that translation occurs solely in the cytoplasm; I test whether some occurs in nuclei and find: (1) L-azidohomoalanine (Aha) – a methionine analogue (detected by microscopy after attaching a fluorescent tag using ‘click’ chemistry) – is incorporated within 5 s into nuclei in a process sensitive to the translation inhibitor, anisomycin. (2) Puromycin – another inhibitor that end-labels nascent peptides (detected by immuno-fluorescence) – is similarly incorporated in a manner sensitive to a transcriptional inhibitor. (3) CD2 – a non-nuclear protein – is found in nuclei close to the nascent RNA that encodes it (detected by combining indirect immuno-labelling with RNA fluorescence in situ hybridization using intronic probes); faulty (nascent) RNA is destroyed by a quality-control mechanism sensitive to translational inhibitors. I conclude that substantial translation occurs in the nucleus, with some being closely coupled to transcription and the associated proof-reading. Moreover, most peptides made in both the nucleus and cytoplasm are degraded soon after they are made with half-lives of about one minute.</p> <p>I also collaborated on two additional projects: the purification of mega-complexes (transcription 'factories’) containing RNA polymerases I, II, or III (I used immuno-fluorescence to confirm that each contained the expected constituents), and the demonstration that some ‘factories’ specialize in transcribing genes responding to tumour necrosis factor α – a cytokine that signals through NFκB (I used RNA fluorescence in situ hybridization coupled with immuno-labelling to show active NFκB is found in factories transcribing responsive genes).</p> |
| spellingShingle | Synthetic organic chemistry Protein chemistry Cell Biology Microscopy Biology Baboo, S Nuclear translation |
| title | Nuclear translation |
| title_full | Nuclear translation |
| title_fullStr | Nuclear translation |
| title_full_unstemmed | Nuclear translation |
| title_short | Nuclear translation |
| title_sort | nuclear translation |
| topic | Synthetic organic chemistry Protein chemistry Cell Biology Microscopy Biology |
| work_keys_str_mv | AT baboos nucleartranslation |