Skip to content
1887

Microbiology Society logo Microbiology

Volume 146, Issue 12

PkwA, a WD-repeat protein, is expressed in spore-derived mycelium of and phosphorylation of its WD domain could act as a molecular switch Free

Abstract

WD-repeat proteins are found in all eukaryotes and are implicated in a variety of regulatory functions as a result of protein–protein interactions. PkwA from CCM3352 is a first potential example of a WD-repeat protein in a prokaryotic actinomycete. A mAb (3G2) was generated against the carboxy terminus of PkwA and was used to analyse the expression of PkwA in . PkwA was detected in exponential growth phase following inoculation with spores, but could not be found at any stage of growth following inoculation with vegetative mycelium. PkwA and its WD domain were expressed in as His-tag derivatives and purified on a Talon metal affinity matrix. The WD domain was phosphorylated by Pkg2, a membrane-spanning protein Ser/Thr kinase from ‘’. A membrane fraction from an exponential, spore-derived culture of was found to phosphorylate the WD domain specifically in the presence of Mn. These data confirm that PkwA is expressed in spore-derived exponential growth phase of and could play a role as a molecular switch in a signalling pathway.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): DIG, digoxigenin , MBP, maltose-binding protein , phosphorylation , PkwA , protein Ser/Thr kinase , Thermomonospora curvata , WD, Trp-Asp , WD-repeat protein and WDA, WD domain of PkwA
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-146-12-3259
2000-12-01
2025-04-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/146/12/1463259a.html?itemId=/content/journal/micro/10.1099/00221287-146-12-3259&mimeType=html&fmt=ahah

References

  1. Bhalerao R. P., Salchert K., Bako L., Okresz L., Szabados L., Muranaka T., Machida Y., Schell J., Koncz C. 1999; Regulatory interaction of PRL1 WD protein with Arabidopsis SNF1-like protein kinases. Proc Natl Acad Sci USA 96:5322–5327 [CrossRef]
     [Google Scholar]
  2. Bormann E. R., Eikmanns B. J., Sahm H. 1992; Molecular analysis of the Corynebacterium glutamicum gdh gene encoding glutamate dehydrogenase. Mol Microbiol 6:317–326 [CrossRef]
     [Google Scholar]
  3. Claney C. E., Mendoza M. G., Naismith T. V., Komnan M. F., Egelhoff T. T. 1997; Identification of a protein kinase from Dictyostelium with homology to the novel catalytic domain of myosin heavy chain kinase A. J Biol Chem 272:11812–11815 [CrossRef]
     [Google Scholar]
  4. Datta P. K., Chytil A., Gorska A. E., Moses H. L. 1998; Identification of STRAP, a novel WD domain protein in transforming growth factor-beta signaling. J Biol Chem 273:34671–34674 [CrossRef]
     [Google Scholar]
  5. Fong H. K., Hurley J. B., Hopkins R. S., Miake-Lye R., Johnson M. S., Doolittle R. F., Simon M. I. 1986; Repetitive segmental structure of the transducin beta subunit: homology with the CDC4 gene and identification of related mRNAs. Proc Natl Acad Sci USA 83:2162–2166 [CrossRef]
     [Google Scholar]
  6. Futey L. M., Medley Q. G., Cote G. P., Egelhoff T. T. 1995; Structural analysis of myosin heavy chain kinase A from Dictyostelium. J Biol Chem 270:523–529 [CrossRef]
     [Google Scholar]
  7. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. 1977; Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature 266:550 [CrossRef]
     [Google Scholar]
  8. Harris J. F., Hawley R. G., Hawley T. S., Crawford-Sharpe G. C. 1992; Increased frequency of both total and specific monoclonal antibody producing hybridomas using a fusion partner that constitutively expresses recombinant IL-6. J Immunol Methods 148:199–207 [CrossRef]
     [Google Scholar]
  9. Hoecker U., Tepperman J. M., Quail P. H. 1999; SPA1, a WD-repeat protein specific to phytochrome A signal transduction. Science 284:496–499 [CrossRef]
     [Google Scholar]
  10. Janda L., Tichy P., Spizek J., Petricek M. 1996; A deduced Thermomonospora curvata protein containing serine/threonine protein kinase and WD-repeat domains. J Bacteriol 178:1487–1489
     [Google Scholar]
  11. Janda L., Pavelka T., Tichy P., Spizek J., Petricek M. 1997; Production and properties of α-glucosidase from the thermotolerant bacterium Thermomonospora curvata. J Appl Microbiol 83:470–476 [CrossRef]
     [Google Scholar]
  12. Johnson L. N., Noble E. M., Owen D. J. 1996; Active and inactive protein kinases: structural basis for regulation. Cell 85:149–158 [CrossRef]
     [Google Scholar]
  13. Kolman M. F., Egelhoff T. T. 1997; Dictyostelium myosin heavy chain kinase A subdomains. Coiled-coil and WD repeat roles in oligomerization and substrate targeting. J Biol Chem 272:16904–16910 [CrossRef]
     [Google Scholar]
  14. Kwak J. M., Kim S. A., Lee S. K., Oh S. A., Byoun C. H., Han J. K., Nam H. G. 1997; Insulin-induced maturation of Xenopus oocytes is inhibited by microinjection of a Brassica napus cDNA clone with high similarity to a mammalian receptor for activated protein kinase C. Planta 201:245–251 [CrossRef]
     [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685 [CrossRef]
     [Google Scholar]
  16. Mannai M., Cozzone A. J. 1982; Endogenous protein phosphorylation in Escherichia coli extracts. Biochem Biophys Res Commun 107:981–988 [CrossRef]
     [Google Scholar]
  17. Matsumoto A., Hong S.-K., Ishizuka H., Horinouchi S., Beppu T. 1994; Phosphorylation of the AfsR protein involved in secondary metabolism in Streptomyces species by a eukaryotic-type protein kinase. Gene 146:47–56 [CrossRef]
     [Google Scholar]
  18. Nadvornik R., Vomastek T., Janecek J., Technikova Z., Branny P. 1999; Pkg2, a novel transmembrane protein Ser/Thr kinase of Streptomyces granaticolor. J Bacteriol 181:15–23
     [Google Scholar]
  19. Neer E. J., Smith T. F. 1996; G protein heterodimers: new structures propel new questions. Cell 84:175–178 [CrossRef]
     [Google Scholar]
  20. Neer E. J., Schmidt C. J., Nambudripad R., Smith T. F. 1994; The ancient regulatory-protein family of WD-repeat proteins. Nature 371:297–300 [CrossRef]
     [Google Scholar]
  21. Peirs P., De Wit L., Braibant M., Huygen K., Content J. 1997; A serine/threonine protein kinase from Mycobacterium tuberculosis. Eur J Biochem 244:604–612 [CrossRef]
     [Google Scholar]
  22. Petricek M., Stajner K., Tichy P. 1989; Cloning of a thermostable α-amylase gene from Thermomonospora curvata and its expression in Streptomyces lividans. J Gen Microbiol 135:3303–3309
     [Google Scholar]
  23. Petricek M., Tichy P., Kuncova M. 1992; Characterization of the α-amylase-encoding gene from Thermomonospora curvata. Gene 112:77–83 [CrossRef]
     [Google Scholar]
  24. Ron D., Chen C.-H., Caldwell J., Jamieson L., Orr E., Mochly-Rosen D. 1994; Cloning of an intracellular receptor for protein kinase C: a homolog of the β subunit of G proteins. Proc Natl Acad Sci USA 91:839–843 [CrossRef]
     [Google Scholar]
  25. Ron D., Luo J., Mochly-Rosen D. 1995; C2 region-derived peptides inhibit translocation and function of β protein kinase C in vivo. J Biol Chem 270:24180–24187 [CrossRef]
     [Google Scholar]
  26. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  27. Sheaff R. J., Groudine M., Gordon M., Roberts J. M., Clurman B. E. 1997; Cyclin E-CKD2 is a regulator of p27Kip1. Genes Dev 11:1464–1478 [CrossRef]
     [Google Scholar]
  28. Smith T. F., Gaitatzes C., Suxena K., Neer E. J. 1999; The WD repeat: a common architecture for diverse functions. Trends Biochem Sci 24:181–185 [CrossRef]
     [Google Scholar]
  29. Udo H., Inouye M., Inouye S. 1997; Biochemical characterization of Pkn2, a protein serine/threonine kinase from Myxococcus xanthus, a Gram-negative developmental bacterium. FEBS Lett 400:188–192 [CrossRef]
     [Google Scholar]
  30. Urabe H., Ogawara H. 1995; Cloning, sequencing and expression of serine/threonine kinase-encoding genes from Streptomyces coelicolor A3(2). Gene 153:99–104 [CrossRef]
     [Google Scholar]
  31. Vomastek T., Nadvornik R., Janecek J., Technikova Z., Weiser J., Branny P. 1998; Characterisation of two putative protein Ser/Thr kinases from actinomycete Streptomyces granaticolor both endowed with different properties. Eur J Biochem 257:55–61 [CrossRef]
     [Google Scholar]
  32. Yam C. H., Ng R. W. M., Siu W. Y., Lau A. W. S., Poon R. Y. C. 1999; Regulation of cyclin A-Cdk2 by SCF component Skp1 and F-box protein Skp2. Mol Cell Biol 19:635–645
     [Google Scholar]
  33. Zhang C.-C. 1996; Bacterial signalling involving eukaryotic-type protein kinases. Mol Microbiol 20:9–15 [CrossRef]
     [Google Scholar]
  34. Zhang H., Hannon G. H., Beach D. 1994; p21-containing cycline kinases exist in both active and inactive states. Genes Dev 8:1750–1758 [CrossRef]
     [Google Scholar]
/content/journal/micro/10.1099/00221287-146-12-3259
Loading
PkwA, a WD-repeat protein, is expressed in spore-derived mycelium of Thermomonospora curvata and phosphorylation of its WD domain could act as a molecular switch
Microbiology 146, 3259 (2000); https://doi.org/10.1099/00221287-146-12-3259
/content/journal/micro/10.1099/00221287-146-12-3259
/content/journal/micro/10.1099/00221287-146-12-3259
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error