- W.R. Hagen (1)
- C. Hetke (1)
- E. Heus de (1)
- S.W.M. Kengen (1)
- T. Lammers (1)
- J.H.G. Lebbink (1)
- J. Oost van der (3)
- G. Schut (2)
- M. Thomm (4)
- J.E. Tuininga (1)
- W.G.B. Voorhorst (1)
- W.M. Vos de (4)
The transcriptional regulator LrpA from the hyperthermophilic archaeon Pyrococcus furiosus is negatively autoregulated
Brinkman, A.B. ; Dahlke, I. ; Tuininga, J.E. ; Lammers, T. ; Dumay, V. ; Heus, E. de; Lebbink, J.H.G. ; Thomm, M. ; Vos, W.M. de; Oost, J. van der - \ 2000
Journal of Biological Chemistry 275 (2000). - ISSN 0021-9258 - p. 38160 - 38169.
The archaeal transcriptional initiation machinery closely resembles core elements of the eukaryal polymerase II system. However, apart from the established basal archaeal transcription system, little is known about the modulation of gene expression in archaea. At present, no obvious eukaryal-like transcriptional regulators have been identified in archaea. Instead, we have previously isolated an archaeal gene, the Pyrococcus furiosus lrpA, that potentially encodes a bacterial-like transcriptional regulator. In the present study, we have for the first time addressed the actual involvement of an archaeal Lrp homologue in transcription modulation. For that purpose, we have produced LrpA in Escherichia coli. In a cell-free P. furiosus transcription system we used wild-type and mutated lrpA promoter fragments to demonstrate that the purified LrpA negatively regulates its own transcription. In addition, gel retardation analyses revealed a single protein-DNA complex, in which LrpA appeared to be present in (at least) a tetrameric conformation. The location of the LrpA binding site was further identified by DNaseI and hydroxyl radical footprinting, indicating that LrpA binds to a 46-base pair sequence that overlaps the transcriptional start site of its own promoter. The molecular basis of the transcription inhibition by LrpA is discussed.
|Transcriptional regulation in the hyperthermophilic archaeon Pyrococcus furiosus: Coordinated expression of divergently oriented genes in response to ß-linked glucose polymers
Voorhorst, W.G.B. ; Gueguen, Y. ; Geerling, A.C.M. ; Schut, G. ; Dahlke, I. ; Thomm, M. ; Oost, J. van der; Vos, W.M. de - \ 1999
Journal of Bacteriology 181 (1999)12. - ISSN 0021-9193 - p. 3777 - 3783.
The ferredoxin-dependent conversion of glyceraldehyde-3-phosphate in the hyperthermophilic archaeon Pyrococcus furiosus represents a novel site of glycolytic regulation.
Oost, J. van der; Schut, G. ; Kengen, S.W.M. ; Hagen, W.R. ; Thomm, M. ; Vos, W.M. de - \ 1998
Journal of Biological Chemistry 273 (1998). - ISSN 0021-9258 - p. 28149 - 28154.
|A cell-free transcription system for the hyperthermophilic archeaon Pyrococcus furiosus.
Hetke, C. ; Geerling, A.C.M. ; Vos, W.M. de; Thomm, M. - \ 1996
Nucleic Acids Research 24 (1996). - ISSN 0305-1048 - p. 2369 - 2376.