Abstract
The formylmethanofuran:tetrahydromethanopterin formyltransferase (FTR) from Methanothermus fervidus was partially purified and its N-terminal amino acid sequence determined. Using as probe a mixture of oligonucleotides derived from the FTR N-terminus, the corresponding gene (ftr) was cloned and sequenced. The ftr gene codes for 297 amino acids, corresponding to a molecular mass of 31836 daltons, in contrast to the 41000 daltons estimated for the protein by sodium dodecylsulphate-polyacrylamide gel electrophoresis. The deduced amino acid sequence of the hyperthermophilic FTR from M. fervidus is 76% identical to the thermophilic FTR from Methanobacterium thermoautotrophicum and has a larger number of lysine residues. A putative ATP-binding site of the FTR is reported. The size of the ftr mRNA was estimated as 1000 nucleotides indicating monocistronic transcription of the 891 by gene. The ftr mRNA starts 27 by downstream of the centre of a putative archaeal box A motif and terminates at an oligo-dT stretch. In vitro transcription of the ftr gene, utilizing a transcription system developed for the distantly related Sulfolobus shibatae, is discussed with respect to the functional conservation of the basal transcription apparatus of Archaea.
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References
Brown JW, Daniels CJ, Reeve IN (1989) Gene structure, organization, and expression in archaebacteria. CRC Crit Rev Microbiol 16:287–338
Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299
DiMarco AA, Sment KA, Konisky J, Wolfe RS (1990) The formylmethanofuran: tetrahydromethanopterin formyltransferase from Methanobacterium thermoautotrophicum DH: Nucleotide sequence and functional expression of the cloned gene. J Biol Chem 265:472–476
Donnelly MI, Wolfe RS (1986) The role of formylmethanofuran: tetrahydromethanopterin formyltransferase in methanogenesis from carbon dioxide. J Biol Chem 261:16653–16659
Eckerskorn C, Mewes W, Goretzki H, Lottspeich F (1988) A new siliconized-glass fiber as support for protein-chemical analysis of electroblotted proteins. Eur J Biochem 176: 509–519
Fabry S, Hensel R (1988) Primary structure of glyceraldehyde-3-phosphate dehydrogenase deduced from the nucleotide sequence of the thermophilic archaebacterium Methanothermus fervidus. Gene 64:189–197
Fabry S, Lang J, Niermann T, Vingron M, Hensel R (1989) Nucleotide sequence of the glyceraldehyde-3-phosphate dehydrogenase gene from the mesophilic methanogenic archaebacteria Methanobacterium bryantii and Methanobacterium formicicum: Comparison with the respective gene structure of the closely related extreme thermophile Methanothermus fervidus. Eur J Biochem 179:405–413
Fabry S, Heppner P, Dietmaier W, Hensel R (1990) Cloning and sequencing the gene encoding 3-phosphoglycerate kinase from mesophilic Methanobacterium bryantii and thermophilic Methanothermus fervidus. Gene 91:19–25
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Haas ES, Brown JW, Daniels CJ, Reeve IN (1990) Genes encoding the 7S RNA and tRNASer are linked to one of the two rRNA operons in the genome of the extremely thermophilic archaebacterium Methanothermus fervidus. Gene 90:51–59
Hain J, Reiter WD, Hudepohl U, Zillig W (1992) Elements of an archaeal promoter defined by mutational analysis. Nucleic Acids Res 20:5423–5428
Hausner W, Frey G, Thomm M (1991) Control regions of an archaeal gene. A TATA box and an initiator element promote cell-free transcription of the tRNAVal gene of Methanococcus vannieli. J Mol Biol 222:495–508
Hensel R, König H (1988) Thermoadaptation of methanogenic bacteria by intracellular ion concentration. FEMS Microbiol Lett 49:75–79
Hüdepohl U, Reiter WD, Zillig W (1990) In vitro transcription of two rRNA genes of the archaebacterium Sulfolobus B12 indicates a factor requirement for specific initiation. Proc Natl Acad Sci USA 87:5851–5855
Hüdepohl U, Gropp F, Horne M, Zillig W (1991) Heterologous in vitro transcription from two archaebacterial promoters. FEBS Lett 285:257–259
Kabsch W, Sander C (1983) Dictionary of protein secondary structure: Pattern recognition of hydrogen-bond and geometrical features. Biopolymers 22:2577–2637
Knaub S, Klein A (1990) Specific transcription of cloned Methanobacterium thermoautotrophicum transcription units by homologous RNA polymerase in vitro. Nucleic Acids Res 18:1441–1446
Lehmacher A, Vogt AB, Hensel R (1990) Biosynthesis of cyclic 2,3-diphosphoglycerate: Isolation and characterization of 2-phosphoglycerate kinase and cyclic 2,3-diphosphoglycerate synthetase from Methanothermus fervidus. FEBS Lett 272:94–98
Menendez-Arias L, Argos P (1989) Engineering protein thermal stability. J Mol Biol 206:397–406
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Saraste M, Sibbald PR, Wittinghofer A (1990) The P-loop — a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci 15:430–434
Selden RF (1989) Analysis of RNA by Northern hybridization. In: Ausubel F, Brent R, Kingston R, Moore D, Seidman J, Smith JA, Struhl K (eds) Current protocols in molecular biology. Wiley-Interscience, New York, pp 4.9.1–4.9.8
Steigerwald VJ, Beckler GS, Reeve JN (1990) Conservation of hydrogenase and polyferrodoxin structures in the hyperthermophilic archaebacterium Methanothermus fervidus. J Bacteriol 172:4715–4718
Stetter KO, Thomm M, Winter J, Wildgruber G, Huber H, Zillig W, Janécovic D, König H, Palm P, Wunderl S (1981) Methanothermus fervidus, sp. nov., a novel extremely thermophilic methanogen isolated from icelandic hot springs. Zentralbl Bakteriol Hyg l. Abt Orig C 2:166–178
Thomm M, Sandman K, Frey G, Koller G, Reeve JN (1992) Transcription in vivo and in vitro of the histone-encoding gene hmfB from the hyperthermophilic archaeon Methanothermus fervidus. J Bacteriol 174:3508–3513
Watson HC, Walker NP, Shaw PJ, Bryant TN, Wendell PL, Fothergill LA, Perkins RE, Conroy SC, Dobson MJ, Tuite MF, Kingsman AJ, Kingsman SM (1982) Sequence and structure of yeast phosphoglycerate kinase. EMBO J 1:1635–1640
Weil CF, Cram DS, Sherf BA, Reeve JN (1988) Structure and comparative analysis of the genes encoding component C of methyl coenzyme M reductase in the extremely thermophilic archaebacterium Methanothermus fervidus. J Bacteriol 170:4718–4726
Zuber H (1988) Temperature adaptation of lactate dehydrogenase: structural, functional and genetic aspects. Biophys Chem 29:171–179
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Lehmacher, A. Cloning, sequencing and transcript analysis of the gene encoding formylmethanofuran: tetrahydromethanopterin formyltransferase from the hyperthermophilic Methanothermus fervidus . Molec. Gen. Genet. 242, 73–80 (1994). https://doi.org/10.1007/BF00277350
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DOI: https://doi.org/10.1007/BF00277350