Nucleic Acids Research, 1990, Vol. 18, No. 6 1361-1367
© 1990
MOLECULAR BIOLOGY |
An archaebacterial cell-free transcription system. The expression of tRNA genes from Methanococcus vannielii is mediated by a transcription factor
Lehrstuhl für Mikrobiologie, Universität Regensburg, Universitätsstr 31 D-8400 Regensburg, FRG
*To whom correspondence should be addressed
Received January 19, 1990. Accepted February 20, 1990.
Our understanding of the mechanism of RNA biosynthesis in archaebacteria is limited, due in part to the inability of purified RNA polymerases to transcribe purified genes accurately in vitro. In the present study, we show that cell extracts of Methanococcus vannielli and Methanococcus thermolithotrophicus purified by gradient centrifugation synthesize a distinct transcript from templates harboring a cloned homologous tRNAVal and tRNAArg gene. The in vitro transcripts initiate with GTP at the same sites as in Methanococcus cells. About 60% of the sequence of the in vitro RNA products was analyzed by dideoxyterminated primer extension and found to be Identical with that of the precursors of tRNAVal and tRNAArg This findings Indicate that this RNA polymerase fraction both initiates and terminates transcription faithfully in vitro.
After purification of a cell-free extract (S-100) of M. thermolithotrophicus by phosphocellulose chromatography, the endogenous RNA polymerase has lost its ability to transcribe the tRNAVal gene accurately. The activity directing specific expression of this template was reconstituted by the addition of a protein-fraction devoid of RNA polymerase activity. Thus, a transcription factor appears to be required for accurate cell-free expression of tRNA genes from M. vannlelii.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Kessler, G. Sezonov, J. I. Guijarro, N. Desnoues, T. Rose, M. Delepierre, S. D. Bell, and D. Prangishvili A novel archaeal regulatory protein, Sta1, activates transcription from viral promoters Nucleic Acids Res., October 18, 2006; 34(17): 4837 - 4845. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Ouhammouch and E. P. Geiduschek An expanding family of archaeal transcriptional activators PNAS, October 25, 2005; 102(43): 15423 - 15428. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. S. Bartlett, M. Thomm, and E. P. Geiduschek Topography of the Euryarchaeal Transcription Initiation Complex J. Biol. Chem., February 13, 2004; 279(7): 5894 - 5903. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Vierke, A. Engelmann, C. Hebbeln, and M. Thomm A Novel Archaeal Transcriptional Regulator of Heat Shock Response J. Biol. Chem., January 3, 2003; 278(1): 18 - 26. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. Dahlke and M. Thomm A Pyrococcus homolog of the leucine-responsive regulatory protein, LrpA, inhibits transcription by abrogating RNA polymerase recruitment Nucleic Acids Res., February 1, 2002; 30(3): 701 - 710. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 W. Hausner and M. Thomm Events during Initiation of Archaeal Transcription: Open Complex Formation and DNA-Protein Interactions J. Bacteriol., May 15, 2001; 183(10): 3025 - 3031. [Abstract] [Full Text]
|
|
|
|
|
|
W. Hausner, U. Lange, and M. Musfeldt Transcription Factor S, a Cleavage Induction Factor of the Archaeal RNA Polymerase J. Biol. Chem., April 21, 2000; 275(17): 12393 - 12399. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Hethke, A. Bergerat, W. Hausner, P. Forterre, and M. Thomm Cell-Free Transcription at 95{degrees}: Thermostability of Transcriptional Components and DNA Topology Requirements of Pyrococcus Transcription Genetics, August 1, 1999; 152(4): 1325 - 1333. [Abstract] [Full Text]
|
|
|
|
|
|
W. G. B. Voorhorst, Y. Gueguen, A. C. M. Geerling, G. Schut, I. Dahlke, M. Thomm, J. van der Oost, and W. M. de Vos Transcriptional Regulation in the Hyperthermophilic Archaeon Pyrococcus furiosus: Coordinated Expression of Divergently Oriented Genes in Response to beta -Linked Glucose Polymers J. Bacteriol., June 15, 1999; 181(12): 3777 - 3783. [Abstract] [Full Text]
|
|
|
|
|
|
W. Hausner, J. Wettach, C. Hethke, and M. Thomm Two Transcription Factors Related with the Eucaryal Transcription Factors TATA-binding Protein and Transcription Factor IIB Direct Promoter Recognition by an Archaeal RNA Polymerase J. Biol. Chem., November 22, 1996; 271(47): 30144 - 30148. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Hausner and M. Thomm The Translation Product of the Presumptive Thermococcus celer TATA-binding Protein Sequence Is a Transcription Factor Related in Structure and Function to Methanococcus Transcription Factor B J. Biol. Chem., July 28, 1995; 270(30): 17649 - 17651. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. B. Brinkman, I. Dahlke, J. E. Tuininga, T. Lammers, V. Dumay, E. de Heus, J. H. G. Lebbink, M. Thomm, W. M. de Vos, and J. van der Oost An Lrp-like Transcriptional Regulator from the Archaeon Pyrococcus furiosus Is Negatively Autoregulated J. Biol. Chem., December 1, 2000; 275(49): 38160 - 38169. [Abstract] [Full Text] [PDF]
|
|