title: Archaeal RNA polymerase subunits E and F are not required for transcription in vitro, but a Thermococcus kodakarensis mutant lacking subunit F is temperature-sensitive.
abstract: All archaeal genomes encode RNA polymerase (RNAP) subunits E and F that share a common ancestry with the eukaryotic RNAP subunits A43 and A14 (Pol I), Rpb7 and Rpb4 (Pol II), and C25 and C17 (Pol III). By gene replacement, we have isolated archaeal mutants of Thermococcus kodakarensis with the subunit F-encoding gene (rpoF) deleted, but we were unable to isolate mutants lacking the subunit E-encoding gene (rpoE). Wild-type T. kodakarensis grows at temperatures ranging from 60 degrees C to 100 degrees C, optimally at 85 degrees C, and the DeltarpoF cells grew at the same rate as wild type at 70 degrees C, but much slower and to lower cell densities at 85 degrees C. The abundance of a chaperonin subunit, CpkB, was much reduced in the DeltarpoF strain growing at 85 degrees C and increased expression of cpkB, rpoF or rpoE integrated at a remote site in the genome, using a nutritionally regulated promoter, improved the growth of DeltarpoF cells. RNAP preparations purified from DeltarpoF cells lacked subunit F and also subunit E and a transcription factor TFE that co-purifies with RNAP from wild-type cells, but in vitro, this mutant RNAP exhibited no discernible differences from wild-type RNAP in promoter-dependent transcription, abortive transcript synthesis, transcript elongation or termination.
authors.affiliation: Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, PA 16802, USA.
authors.forename: Akira
authors.initials: A
authors.lastname: Hirata
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Kanai
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Santangelo
authors.forename: Momoko
authors.initials: M
authors.lastname: Tajiri
authors.forename: Kenji
authors.initials: K
authors.lastname: Manabe
authors.forename: John N
authors.initials: JN
authors.lastname: Reeve
authors.forename: Tadayuki
authors.initials: T
authors.lastname: Imanaka
authors.forename: Katsuhiko S
authors.initials: KS
authors.lastname: Murakami
mesh_terms: D019843:Archaeal Proteins / Q000235:genetics* / Q000378:metabolism; D019641:DNA, Archaeal / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000235:genetics* / Q000378:metabolism; D017353:Gene Deletion; D006358:Hot Temperature; D009154:Mutation; D010641:Phenotype; D010957:Plasmids; D012808:Sigma Factor / Q000235:genetics* / Q000378:metabolism; D019712:Thermococcus / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D019843:Archaeal Proteins; D019641:DNA, Archaeal; D012808:Sigma Factor; D012321:DNA-Directed RNA Polymerases
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):6969-73
references.pmid: 12746498
references.citation: J Biol Chem. 2004 Dec 10;279(50):51719-21
references.pmid: 15485836
references.citation: Biochim Biophys Acta. 2002 Sep 10;1555(1-3):83-91
references.pmid: 12206896
references.citation: J Bacteriol. 2001 Mar;183(5):1813-8
references.pmid: 11160119
references.citation: Mol Microbiol. 2004 May;52(4):1133-43
references.pmid: 15130130
references.citation: EMBO J. 2001 Jun 1;20(11):2823-34
references.pmid: 11387215
references.citation: Archaea. 2004 Oct;1(4):263-7
references.pmid: 15810436
references.citation: J Bacteriol. 2003 Jan;185(1):210-20
references.pmid: 12486058
references.citation: J Biol Chem. 2007 Nov 16;282(46):33659-33670
references.pmid: 17875647
references.citation: Mol Cell. 2002 Sep;10(3):635-46
references.pmid: 12408830
references.citation: EMBO Rep. 2001 Feb;2(2):133-8
references.pmid: 11258705
references.citation: EMBO J. 2000 Oct 16;19(20):5473-82
references.pmid: 11032814
references.citation: J Bacteriol. 2008 Jan;190(1):157-67
references.pmid: 17965161
references.citation: Curr Opin Genet Dev. 2004 Apr;14(2):133-8
references.pmid: 15196459
references.citation: Mol Cell Biol. 1999 Apr;19(4):2672-80
references.pmid: 10082533
references.citation: J Biol Chem. 2007 Dec 7;282(49):35482-90
references.pmid: 17921145
references.citation: Biochem Biophys Res Commun. 2001 Jan 19;280(2):581-7
references.pmid: 11162559
references.citation: J Mol Biol. 2006 Jan 13;355(2):196-210
references.pmid: 16305799
references.citation: Cell. 2007 Dec 28;131(7):1260-72
references.pmid: 18160037
references.citation: Trends Microbiol. 1998 Jun;6(6):222-8
references.pmid: 9675798
references.citation: J Biol Chem. 2001 Dec 7;276(49):46408-13
references.pmid: 11577101
references.citation: Mol Cell Biol. 1993 Nov;13(11):6984-91
references.pmid: 8413288
references.citation: J Bacteriol. 2007 Apr;189(7):2683-91
references.pmid: 17259314
references.citation: Mol Cell. 2003 Oct;12(4):1003-13
references.pmid: 14580350
references.citation: Yeast. 1993 Mar;9(3):295-9
references.pmid: 8488730
references.citation: Appl Environ Microbiol. 2008 May;74(10):3099-104
references.pmid: 18378640
references.citation: Mol Cell. 2006 Jul 7;23(1):71-81
references.pmid: 16818233
references.citation: J Biol Chem. 2001 Mar 30;276(13):10097-102
references.pmid: 11087726
references.citation: Mol Microbiol. 2007 Sep;65(6):1395-404
references.pmid: 17697097
references.citation: Genome Res. 2005 Mar;15(3):352-63
references.pmid: 15710748
references.citation: Appl Environ Microbiol. 2005 Jul;71(7):3889-99
references.pmid: 16000802
references.citation: J Mol Biol. 2007 Mar 23;367(2):344-57
references.pmid: 17275836
references.citation: Mol Cell. 2001 Nov;8(5):1137-43
references.pmid: 11741548
references.citation: J Biol Chem. 2003 Jan 31;278(5):3339-46
references.pmid: 12429747
references.citation: Trends Genet. 2008 May;24(5):211-5
references.pmid: 18384908
references.citation: Proc Natl Acad Sci U S A. 2003 Jun 10;100(12):6964-8
references.pmid: 12746495
references.citation: Mol Cell. 2007 Mar 23;25(6):813-23
references.pmid: 17386259
references.citation: J Biol Chem. 1991 Jan 5;266(1):71-5
references.pmid: 1985924
references.citation: Mol Microbiol. 2000 Feb;35(3):667-76
references.pmid: 10672188
references.citation: Mol Cell Biol. 1989 Jul;9(7):2854-9
references.pmid: 2674672
references.citation: Nature. 2008 Feb 14;451(7180):851-4
references.pmid: 18235446
references.citation: J Biol Chem. 2007 Apr 13;282(15):11047-57
references.pmid: 17311916