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Functional analysis of multiple general transcription factors in Sulfolobus acidocaldarius ; Funktionsanalyse multipler genereller Transkriptionsfaktoren in Sulfolobus acidocaldarius
The archaeal transcription machinery encompasses one multi-subunit RNA polymerase (RNAP), resembling the RNAPII of Eukaryotes, homologs of the TATA-binding protein (TBP), transcription factor TFIIB (TFB) and TFIIE-α (TFE) (Langer and Zillig, 1993, Marsh et al., 1994, Langer et al., 1995, Kyrpides and Ouzounis 1999, Bell and Jackson 1998a, b, Bell and Jackson, 2001, Hickey et al., 2002, Geiduscheck and Ouhammouch, 2005, Werner and Grohmann, 2011). The current mechanistic understanding of transcription initiation is that TBP binds to the TATA-box (~25 bp upstream of the transcription start site) whereupon TFB binds to the TBP-DNA complex forming sequence specific contacts with a purine-rich TFB-responsive element (BRE). Subsequently, the N-terminus of TFB recruits the RNAP to build the ternary pre-initiation complex. RNAP, TBP and TFB are solely sufficient for transcription of archaeal promoters in vitro (Bell and Jackson 1998a, b, Bell and Jackson, 2001, Hickey et al., 2002, Geiduscheck and Ouhammouch, 2005). Therefore archaeal transcription is generally regarded as a simpler model of the more complex eukaryal processes (Baliga et al. 2000). Whereas multiple tfb and tbp homologs have been deeply investigated in the euryarchaeal branch, the role of multiple GTFs in Crenarchaeota remains poorly understood. It has been proposed that various tfb homologs might serve like σ-factors in Bacteria and fulfill specialized functions according to stress response (Micorescu et al., 2008, Peng et al., 2009, Paytubi and White, 2009). The aim of this study was to provide multiple GTF proteins required for experiments like EMSAs and in vitro transcription and to gain a deeper insight into their function in S. acidocaldarius by the generation of reporter gene constructs, protein-protein interaction studies and generation of knock-out mutants. The thermoacidophile S. acidocaldarius grows optimally at 80°C and pH 2-3 (Brock et al., 1972) and is an obligate aerobic organism; it grows heterotrophically and requires a limited range of ...
Functional analysis of multiple general transcription factors in Sulfolobus acidocaldarius ; Funktionsanalyse multipler genereller Transkriptionsfaktoren in Sulfolobus acidocaldarius
The archaeal transcription machinery encompasses one multi-subunit RNA polymerase (RNAP), resembling the RNAPII of Eukaryotes, homologs of the TATA-binding protein (TBP), transcription factor TFIIB (TFB) and TFIIE-α (TFE) (Langer and Zillig, 1993, Marsh et al., 1994, Langer et al., 1995, Kyrpides and Ouzounis 1999, Bell and Jackson 1998a, b, Bell and Jackson, 2001, Hickey et al., 2002, Geiduscheck and Ouhammouch, 2005, Werner and Grohmann, 2011). The current mechanistic understanding of transcription initiation is that TBP binds to the TATA-box (~25 bp upstream of the transcription start site) whereupon TFB binds to the TBP-DNA complex forming sequence specific contacts with a purine-rich TFB-responsive element (BRE). Subsequently, the N-terminus of TFB recruits the RNAP to build the ternary pre-initiation complex. RNAP, TBP and TFB are solely sufficient for transcription of archaeal promoters in vitro (Bell and Jackson 1998a, b, Bell and Jackson, 2001, Hickey et al., 2002, Geiduscheck and Ouhammouch, 2005). Therefore archaeal transcription is generally regarded as a simpler model of the more complex eukaryal processes (Baliga et al. 2000). Whereas multiple tfb and tbp homologs have been deeply investigated in the euryarchaeal branch, the role of multiple GTFs in Crenarchaeota remains poorly understood. It has been proposed that various tfb homologs might serve like σ-factors in Bacteria and fulfill specialized functions according to stress response (Micorescu et al., 2008, Peng et al., 2009, Paytubi and White, 2009). The aim of this study was to provide multiple GTF proteins required for experiments like EMSAs and in vitro transcription and to gain a deeper insight into their function in S. acidocaldarius by the generation of reporter gene constructs, protein-protein interaction studies and generation of knock-out mutants. The thermoacidophile S. acidocaldarius grows optimally at 80°C and pH 2-3 (Brock et al., 1972) and is an obligate aerobic organism; it grows heterotrophically and requires a limited range of ...
Functional analysis of multiple general transcription factors in Sulfolobus acidocaldarius ; Funktionsanalyse multipler genereller Transkriptionsfaktoren in Sulfolobus acidocaldarius
Rauch, Bernadette (author) / Siebers, Bettina
2013-12-18
Theses
Electronic Resource
English
Polyphosphate metabolism and profiling of myristoylation in Sulfolobus acidocaldarius
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