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Identifizierung des zentralen Polypeptids des generellen RNA-Polymerase III Transkriptionsfaktors TFIIIC1
Der Transkriptionsfaktor TFIIIC1 wird für die RNA-Synthese aller humanen Polymerase III-abhängigen Gene unabhängig vom jeweiligen Promotortyp essentiell benötigt. Innerhalb des humanen Pol III Systems ist TFIIIC1 der einzige Transkriptionsfaktor dessen Untereinheiten-Zusammensetzung bisher noch imme...
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| Format: | Dissertation |
| Sprache: | Deutsch |
| Veröffentlicht: |
Philipps-Universität Marburg
2004
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| Online Zugang: | PDF-Volltext |
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The human transcription factor TFIIIIC1 is a general factor for transcription of all RNA pol III dependent genes. There are no information about the molecular entity of TFIIIC1. The aim of this work was the purification of TFIIIC1 by chromatography in order to analyse the molecular entity of TFIIIC1 by MALDI TOF. The use of anionic exchanger MiniQ resulted in purification and conzentration of TFIIIC1 fractions which was nesessary for further purification of TFIIIC1 by size exclusion chromatography (SEC). SEC resulted in MALDI TOF analysis of seven polypeptides of the most purified TFIIIC1 fraction. Western blot analyses and immundepletion identified the "Transcription Factor-like Nuclear Regulator" (TFNR) as the central polypeptid of TFIIIC1. The 250 kDa protein TFNR shows N-terminal homology to the 150-160 kDa protein hBdp1 (hB'', TFIIIB150). Bdp1 is discribed as an essential pol III transcription factor and the third component of TFIIIB. The identification of the central polypeptid of TFIIIC1 activity proof the function of the 250 kDa form of hBdp1 (TFNR) as essential pol III transcription factor. The N-terminus of hBdp1 is the essential part for transcription activity in vitro.