A 4.2-kb SphI-BamHI fragment of chromosomal DNA from Streptomyces granaticolor was cloned and shown to encode a protein with significant sequence similarity to the eukaryotic protein serine/threonine kinases. It consists of 701 amino acids and in the N-terminal part contains all conserved catalytic domains of protein kinases. The C-terminal domain of Pkg2 contains seven tandem repeats of 11 or 12 amino acids with similarity to the tryptophan-docking motif known to stabilize a symmetrical three-dimensional structure called a propeller structure. The pkg2 gene was overexpressed in Escherichia coli, and the gene product (Pkg2) has been found to be autophosphorylated at serine and threonine residues. The N- and C-terminal parts of Pkg2 are separated with a hydrophobic stretch of 21 amino acids which translocated a PhoA fusion protein into the periplasm. Thus, Pkg2 is the first transmembrane protein serine/threonine kinase described for streptomycetes. Replacement of the pkg2 gene by the spectinomycin resistance gene resulted in changes in the morphology of aerial hyphae.

Cell and Molecular Microbiology Division Institute of Microbiology Czech Academy of Sciences 142 20 Prague 4 Czech Republic

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Altschul S F, Gish W, Miller W, Myers E W, Lipman D J. Basic local alignment search tool. J Mol Biol. 1990;215:403–410. PubMed

Anthony C, Ghosh M, Blake C C. The structure and function of methanol dehydrogenase and related quinoproteins containing pyrrolo-quinolone quinone. Biochem J. 1994;304:665–674. PubMed PMC

Bibb M J, Findlay P R, Johnson M W. The relationship between base composition and codon usage in bacterial genes and its use for the simple and reliable identification of protein-coding sequences. Gene. 1984;30:157–166. PubMed

Blattner F R, Plunkett G R, Bloch C A, Perna N T, Burland V, Riley M, Collado Vides J, Glasner J D, Rode C K, Mayhew G F, Gregor J, Davis N W, Kirkpatrick H A, Goeden M A, Rose D J, Mau B, Shao Y. The complete genome sequence of Escherichia coli K-12. Science. 1997;277:1453–1474. PubMed

Blondelet Rouault M H, Weiser J, Lebrihi A, Branny P, Pernodet J L. Antibiotic resistance gene cassettes derived from the omega interposon for use in E. coli and Streptomyces. Gene. 1997;190:315–317. PubMed

Brickman E, Beckwith J. Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and φ80 transducing phages. J Mol Biol. 1975;96:307–316. PubMed

Carrera A C, Alexandrov K, Roberts T M. The conserved lysine of the catalytic domain of protein kinases is actively involved in the phosphotransfer reaction and not required for anchoring ATP. Proc Natl Acad Sci USA. 1993;90:442–446. PubMed PMC

Chater K F. Taking a genetic scalpel to the Streptomyces colony. Microbiology. 1998;144:1465–1478. PubMed

Cole S T, Brosch R, Parkhill J, Garnier T, Churcher C, Harris D, Gordon S V, Eiglmeier K, Gas S, Barry III C E, Tekaia F, Badcock K, Basham D, Brown D, Chillingworth T, Connor R, Davies R, Devlin K, Feltwell T, Gentles S, Hamlin N, Holroyd S, Hornsby T, Jagels K, Krogh A, McLean J, Moule S, Murphy L, Oliver K, Osborne J, Quail M A, Rajandream M-A, Rogers J, Rutter S, Seeger K, Skelton J, Squares S, Squares R, Sulston J E, Taylor K, Whitehead S, Barrell B G. Deciphering the biology of Mycobacterium tuberculosis from the complete genome sequence. Nature. 1998;393:537–544. PubMed

Cozier G E, Giles I G, Anthony C. The structure of the quinoprotein alcohol dehydrogenase of Acetobacter aceti modelled on that of methanol dehydrogenase from Methylobacterium extorquens. Biochem J. 1995;308:375–379. PubMed PMC

Crennell S J, Garman E F, Laver W G, Vimr E R, Taylor G L. Crystal structure of a bacterial sialidase (from Salmonella typhimurium LT2) shows the same fold as an influenza virus neuraminidase. Proc Natl Acad Sci USA. 1993;90:9852–9856. PubMed PMC

Duclos B, Grangeasse C, Vaganay E, Riberty M, Cozzone A J. Autophosphorylation of a bacterial protein at tyrosine. J Mol Biol. 1996;259:891–895. PubMed

Galyov E E, Hakansson S, Forsberg A, Wolf Watz H. A secreted protein kinase of Yersinia pseudotuberculosis is an indispensable virulence determinant. Nature. 1993;361:730–732. PubMed

Ghosh M, Anthony C, Harlos K, Goodwin M G, Blake C. The refined structure of the quinoprotein methanol dehydrogenase from Methylobacterium extorquens at 1.94 A. Structure. 1995;3:177–187. PubMed

Hanks S K, Quinn A M, Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988;241:42–52. PubMed

Hanlon W A, Inouye M, Inouye S. Pkn9, a Ser/Thr protein kinase involved in the development of Myxococcus xanthus. Mol Microbiol. 1997;23:459–471. PubMed

Hartmann E, Rapoport T A, Lodish H F. Predicting the orientation of eukaryotic membrane-spanning proteins. Proc Natl Acad Sci USA. 1989;86:5786–5790. PubMed PMC

Hoffman C S, Wright A. Fusions of secreted proteins to alkaline phosphatase: an approach for studying protein secretion. Proc Natl Acad Sci USA. 1985;82:5107–5111. PubMed PMC

Hofmann K, Stoffel W. TMbase—a database of membrane spanning protein segments. Biol Chem Hoppe-Seyler. 1993;347:166.

Hopwood D A, Bibb M J, Chater K F, Kieser T, Bruton C J, Kieser H M, Lydiate D J, Smith C P, Ward J M, Schrempf H. Genetic manipulation of Streptomyces. A laboratory manual. Norwich, England: John Innes Foundation; 1985.

Ito N, Phillips S E, Yadav K D, Knowles P F. Crystal structure of a free radical enzyme, galactose oxidase. J Mol Biol. 1994;238:794–814. PubMed

Janda L, Tichý P, Spížek J, Petříček M. A deduced Thermomonospora curvata protein containing serine/threonine protein kinase and WD-repeat domains. J Bacteriol. 1996;178:1487–1489. PubMed PMC

Janeček J, Moravec V, Dobrová Z, Janda I, Weiser J. Protein phosphorylation in submerged spores and vegetative mycelium of Streptomyces granaticolor. FEMS Microbiol Lett. 1995;133:91–94.

Kamps M P, Sefton B M. Acid and base hydrolysis of phosphoproteins bound to Immobilon facilitates analysis of phosphoamino acids in gel-fractionated proteins. Anal Biochem. 1989;176:22–27. PubMed

Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S, Kimura T, Hosouchi T, Matsuno A, Muraki A, Nakazaki N, Naruo K, Okumura S, Shimpo S, Takeuchi C, Wada T, Watanabe A, Yamada M, Yasuda M, Tabata S. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res. 1996;3:109–136. PubMed

Kunkel T A, Roberts J D, Zakour R A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. PubMed

Manai M, Cozzone A J. Endogenous protein phosphorylation in Escherichia coli extracts. Biochem Biophys Res Commun. 1982;107:981–988. PubMed

Manoil C, Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci USA. 1985;82:8129–8133. PubMed PMC

Matsumoto A, Hong S K, Ishizuka H, Horinouchi S, Beppu T. Phosphorylation of the AfsR protein involved in secondary metabolism in Streptomyces species by a eukaryotic-type protein kinase. Gene. 1994;146:47–56. PubMed

Munoz-Dorado J, Inouye S, Inouye M. A gene encoding a protein serine/threonine kinase is required for normal development of M. xanthus, a gram-negative bacterium. Cell. 1991;67:995–1006. PubMed

Oh S H, Chater K F. Denaturation of circular or linear DNA facilitates targeted integrative transformation of Streptomyces coelicolor A3(2): possible relevance to other organisms. J Bacteriol. 1997;179:122–127. PubMed PMC

Pearson W R, Lipman D J. Improved tools for biological sequence comparison. Proc Natl Acad Sci USA. 1988;85:2444–2448. PubMed PMC

Peirs P, DeWit L, Braibant M, Huygen K, Content J. A serine/threonine protein kinase from Mycobacterium tuberculosis. Eur J Biochem. 1997;244:604–612. PubMed

Petříček M, Smrčková I, Tichý P. Transformation of Streptomyces granaticolor with natural and recombinant plasmid vectors. Folia Microbiol. 1985;30:474–478. PubMed

Pope M K, Green B D, Westpheling J. The bld mutants of Streptomyces coelicolor are defective in the regulation of carbon utilization, morphogenesis and cell-cell signalling. Mol Microbiol. 1996;19:747–756. PubMed

Prentki P, Krisch H M. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984;29:303–313. PubMed

Řičicová A, Řeháček Z. Taxonomic characteristic of the strain ETH 7437 producing granaticin. Folia Microbiol. 1968;13:346–349. PubMed

Sambrook J, Fritsch E F, Maniatis T. Molecular cloning: a laboratory manual. 2nd ed. Cold Spring Harbor, N.Y: Cold Spring Harbor Laboratory; 1989.

Sanger F, Nicklen S, Coulson A R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA. 1977;74:5463–5467. PubMed PMC

Stock J B, Ninfa A J, Stock A M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989;53:450–490. PubMed PMC

Studier F W, Rosenberg A H, Dunn J J, Dubendorff J W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. PubMed

Udo H, Munoz-Dorado J, Inouye M, Inouye S. Myxococcus xanthus, a gram-negative bacterium, contains a transmembrane protein serine threonine kinase that blocks the secretion of beta-lactamase by phosphorylation. Genes Dev. 1995;9:972–983. PubMed

Urabe H, Ogawara H. Cloning, sequencing and expression of serine/threonine kinase-encoding genes from Streptomyces coelicolor A3(2) Gene. 1995;153:99–104. PubMed

Varghese J N, Laver W G, Colman P M. Structure of the influenza virus glycoprotein antigen neuraminidase at 2.9 Å resolution. Nature. 1983;303:35–40. PubMed

Vellieux F M, Huitema F, Groendijk H, Kalk K H, Jzn J F, Jongejan J A, Duine J A, Petratos K, Drenth J, Hol W G. Structure of quinoprotein methylamine dehydrogenase at 2.25 Å resolution. EMBO J. 1989;8:2171–2178. PubMed PMC

Vomastek T, Nádvorník R, Janeček J, Techniková Z, Weiser J, Branny P. Characterisation of two putative protein Ser/Thr kinases from actinomycete Streptomyces granaticolor both endowed with different properties. Eur J Biochem. 1998;257:55–61. PubMed

Zhang C C. A gene encoding a protein related to eukaryotic protein kinases from the filamentous heterocystous cyanobacterium Anabaena PCC 7120. Proc Natl Acad Sci USA. 1993;90:11840–11844. PubMed PMC

Zhang C C, Friry A, Peng L. Molecular and genetic analysis of two closely linked genes that encode, respectively, a protein phosphatase 1/2A/2B homolog and a protein kinase homolog in the cyanobacterium Anabaena sp. strain PCC 7120. J Bacteriol. 1998;180:2616–2622. PubMed PMC

Zhang C C, Libs L. Cloning and characterisation of the pknD gene encoding an eukaryotic-type protein kinase in the cyanobacterium Anabaena sp. PCC7120. Mol Gen Genet. 1998;258:26–33. PubMed

Zhang W D, Inouye M, Inouye S. Reciprocal regulation of the differentiation of Myxococcus xanthus by Pkn5 and Pkn6, eukaryotic-like Ser/Thr protein kinases. Mol Microbiol. 1996;20:435–447. PubMed

A eukaryotic-type signalling system of Pseudomonas aeruginosa contributes to oxidative stress resistance, intracellular survival and virulence

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Comparative study of the life cycle dependent post-translation modifications of protein synthesis elongation factor Tu present in the membrane proteome of streptomycetes and mycobacteria

Effect of protein kinase inhibitors on protein phosphorylation and germination of aerial spores from Streptomyces coelicolor

Eukaryotic-type serine/threonine protein kinase StkP is a global regulator of gene expression in Streptococcus pneumoniae

Effect of phosphate on the expression of protein-Ser/Thr kinase pkg2 in Streptomyces granaticolor

Evidence for phosphoprotein phosphatase in Streptomyces granaticolor

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