1887

Microbiology Society logo

Volume 150, Issue 1

Secretion of immunodominant membrane protein from onion yellows phytoplasma through the Sec protein-translocation system in Free

Abstract

A gene that encodes a putative SecE protein, which is a component of the Sec protein-translocation system, was cloned from the onion yellows phytoplasma (OY). The identification of this gene and the previously reported genes encoding SecA and SecY provides evidence that the Sec system exists in phytoplasma. In addition, a gene encoding an antigenic membrane protein (Amp) (a type of immunodominant membrane protein) of OY was cloned and sequenced. The OY gene consisted of 702 nt encoding a protein of 233 aa which was highly similar to Amp of aster yellows phytoplasma (AY). Part of OY Amp was overexpressed in , purified, and used to raise an anti-Amp polyclonal antibody. The anti-Amp antibody reacted specifically with an OY-infected plant extract in Western blot analysis and was therefore useful for the detection of OY as well as Amp. Amp has a conserved protein motif that is known to be exported by the Sec system of . A partial OY Amp protein expressed in was localized in the periplasm as a shorter, putatively processed form of the protein. It had probably been exported from the cytoplasm to the periplasm through the Sec system. Moreover, OY Amp protein expressed in OY and detected in OY-infected plants was apparently also processed. Because phytoplasmas cannot be cultured or transformed, little information is available regarding their protein secretion systems. This study suggests that the Sec system operates in this phytoplasma to export OY Amp.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): Amp, antigenic membrane protein , AP, apple proliferation phytoplasma , AY, aster yellows phytoplasma , CP, clover phyllody phytoplasma , E/H/T, export leader, hydrophilic and transmembrane domains, respectively, in the putative Amp , ESFY, European stone fruit yellows phytoplasma , OY, onion yellows phytoplasma , PD, pear decline phytoplasma , PYLR, peach yellow leaf roll phytoplasma , SPWB, sweet potato witches' broom phytoplasma and WX, western X-disease phytoplasma
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26521-0
2004-01-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/1/mic1500135.html?itemId=/content/journal/micro/10.1099/mic.0.26521-0&mimeType=html&fmt=ahah

References

  1. Akimaru J., Matsuyama S., Tokuda H., Mizushima S. 1991; Reconstitution of a protein translocation system containing purified SecY, SecE, and SecA from Escherichia coli. Proc Natl Acad Sci U S A 88:6545–6549 [CrossRef]
     [Google Scholar]
  2. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J Mol Biol 215:403–410 [CrossRef]
     [Google Scholar]
  3. Altschul S. F., Madden T. L., Schäffer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J. 1997; Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402 [CrossRef]
     [Google Scholar]
  4. Barbara D. J., Morton A., Clark M. F., Davies D. L. 2002; Immunodominant membrane proteins from two phytoplasmas in the aster yellows clade (chlorante aster yellows and clover phyllody) are highly divergent in the major hydrophilic region. Microbiology 148:157–167
     [Google Scholar]
  5. Berg M., Davies D. L., Clark M. F., Vetten H. J., Maier G., Marcone C., Seemuller E. 1999; Isolation of the gene encoding an immunodominant membrane protein of the apple proliferation phytoplasma, and expression and characterization of the gene product. Microbiology 145:1937–1943 [CrossRef]
     [Google Scholar]
  6. Blomquist C. L., Barbara D. J., Davies D. L., Clark M. F., Kirkpatric B. C. 2001; An immunodominant membrane protein gene from the Western X-disease phytoplasma is distinct from those of other phytoplasmas. Microbiology 147:571–580
     [Google Scholar]
  7. Bonnie B., Wilson D. B., Wolf E., Tonchev T., Milla M., Kim P. S. 1995; Predicting coiled coils by use of pairwise residue correlations. Proc Natl Acad Sci U S A 92:8259–8263 [CrossRef]
     [Google Scholar]
  8. Economou A. 1999; Following the leader: bacterial protein export through the Sec pathway. Trends Microbiol 7:315–320 [CrossRef]
     [Google Scholar]
  9. Fraser C. M., Gocayne J. D., White O.26 other authors 1995; The minimal gene complement of Mycoplasma genitalium. Science 270:397–403 [CrossRef]
     [Google Scholar]
  10. Hirokawa T., Seah B.-C., Mitaku S. 1998; SOSUI: classification and secondary structure prediction syst for membrane proteins. Bioinformatics 14:378–379 [CrossRef]
     [Google Scholar]
  11. Jung H.-Y., Sawayanagi T., Kakizawa S. & 7 other authors; 2002; ‘Candidatus Phytoplasma castaneae’, a novel phytoplasma taxon associated with chestnut witches' broom disease. Int J Syst Evol Microbiol 52:1543–1549 [CrossRef]
     [Google Scholar]
  12. Kakizawa S., Oshima K., Kuboyama T. & 7 other authors; 2001; Cloning and expression analysis of phytoplasma protein translocation genes. Mol Plant–Microbe Interact 14:1043–1050 [CrossRef]
     [Google Scholar]
  13. Kirkpatrick B. C. 1992; Mycoplasma-like organisms: plant and invertebrate pathogens. In The Prokaryotes, 2nd edn. pp. 4050–4067Edited by Balows A., Trüper H. G., Dworkin M., Harder W., Schleifer K. H. New York: Springer;
     [Google Scholar]
  14. Lupas A., Van Dyke M., Stock J. 1991; Predicting coiled coils from protein sequences. Science 252:1162–1164 [CrossRef]
     [Google Scholar]
  15. McCoy R. E., Caudwell A., Chang C. J.15 other authors 1989; Plant diseases associated with Mycoplasma-like organisms. In The Mycoplasmas vol 5 pp. 546–640Edited by Whitcomb R. F., Tully J. G. San Diego, CA: Academic Press;
     [Google Scholar]
  16. Medigue C., Wong C.-Y. B., Lin C.-M. M., Bocs S., Danchin A. 2002; The secE gene of Helicobacter pylori. J Bacteriol 184:2837–2840 [CrossRef]
     [Google Scholar]
  17. Morton A., Davies D. L., Blomquist C. L., Barbara D. J. 2003; Characterization of homologues of the apple proliferation immunodominant membrane protein gene from three related phytoplasmas. Mol Plant Pathol 4:109–114 [CrossRef]
     [Google Scholar]
  18. Nakai K., Kanehisa M. 1991; Expert system for predicting protein localization sites in Gram-negative bacteria. Proteins Struct Funct Genet 11:95–110 [CrossRef]
     [Google Scholar]
  19. Nielsen H., Engelbrecht J., Brunak S., von Heijne G. 1997; Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6 [CrossRef]
     [Google Scholar]
  20. Nishiyama K., Suzuki H., Tokuda H. 2000; Role of the non-essential region encompassing the N-terminal two transmembrane stretches of Escherichia coli SecE. Biosci Biotechnol Biochem 64:2121–2127 [CrossRef]
     [Google Scholar]
  21. Oshima K., Miyata S., Sawayanagi T. & 10 other authors; 2002; Minimal set of metabolic pathways suggested from the genome of onion yellows phytoplasma. J Gen Plant Pathol 68:225–236 [CrossRef]
     [Google Scholar]
  22. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  23. Shiomi T., Tanaka M., Wakiya H., Zenbayashi R. 1996; Occurrence of Welsh onion yellows. Ann Phytopathol Soc Jpn 62:258–260 [CrossRef]
     [Google Scholar]
  24. Thompson J. D., Higgins D. G., Gibson T. J. 1994; Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680 [CrossRef]
     [Google Scholar]
  25. Tjalsma H., Bolhuis A., Jongbloed J. D., Bron S., van Dijl J. M. 2000; Signal peptide-dependent protein transport in Bacillus subtilis: a genome-based survey of the secretome. Microbiol Mol Biol Rev 64:515–547 [CrossRef]
     [Google Scholar]
  26. Ueguchi C., Ito K. 1990; Escherichia coli sec mutants accumulate a processed immature form of maltose-binding protein (MBP), a late-phase intermediate in MBP export. J Bacteriol 172:5643–5649
     [Google Scholar]
  27. Voet D., Voet J. 1995 Biochemistry, 2nd edn. pp. 154–155 New York: Wiley;
     [Google Scholar]
  28. Wolfe P. B., Wickner W., Goodman J. M. 1983; Sequence of the leader peptidase gene of Escherichia coli and the orientation of leader peptidase in the bacterial envelope. J Biol Chem 258:12073–12080
     [Google Scholar]
  29. Yu Y.-L., Yeh K.-W., Lin C.-P. 1998; An antigenic protein gene of a phytoplasma associated with sweet potato witches' broom. Microbiology 144:1257–1262 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26521-0
Loading
Secretion of immunodominant membrane protein from onion yellows phytoplasma through the Sec protein-translocation system in Escherichia coli
Microbiology 150, 135 (2004); https://doi.org/10.1099/mic.0.26521-0
/content/journal/micro/10.1099/mic.0.26521-0
/content/journal/micro/10.1099/mic.0.26521-0
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