1887

Abstract

Wsc1 is a member of a family of highly -glycosylated cell surface proteins that reside in the plasma membrane of and function as sensors of cell wall stress. These proteins activate the cell wall integrity signalling pathway by stimulating the small G-protein Rho1, protein kinase C (Pkc1) and a MAP kinase cascade. The cytoplasmic domains of Wsc1 family members interact with the Rom2 guanine nucleotide exchange factor to stimulate GTP-binding of Rho1. Here, a mutational analysis of the cytoplasmic domain of Wsc1 is presented. The data identify two regions of the Wsc1 cytoplasmic tail that are conserved with other family members as important for Rom2 interaction. These regions are separated by an inhibitory region, which includes a cluster of seryl residues that appear to be phosphorylated. Mutational analysis of these residues supports a model in which Wsc1 interaction with Rom2 is negatively regulated by phosphorylation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.27264-0
2004-10-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/10/mic1503281.html?itemId=/content/journal/micro/10.1099/mic.0.27264-0&mimeType=html&fmt=ahah

References

  1. Buehrer, B. & Errede, B.(1997). Coordination of the mating and cell integrity mitogen-activated protein kinase pathways in Saccharomyces cerevisiae. Mol Cell Biol 17, 6517–6525. [Google Scholar]
  2. Cid, V. J., Duran, A., Rey, F., Snyder, M. P., Nombela, C. & Sanchez, M.(1995). Molecular basis of cell integrity and morphogenesis in Saccharomyces cerevisiae. Microbiol Rev 59, 345–386. [Google Scholar]
  3. Costigan, C., Gehrung, S. & Snyder, M.(1992). A synthetic lethal screen identifies SLK1, a novel protein kinase homolog implicated in yeast cell morphogenesis and cell growth. Mol Cell Biol 12, 1162–1178. [Google Scholar]
  4. Davenport, K. R., Sohaskey, M., Kamada, Y., Levin, D. E. & Gustin, M. C.(1995). A second osmosensing signal transduction pathway in yeast. J Biol Chem 270, 30157–30161.[CrossRef] [Google Scholar]
  5. Douglas, C. M., Foor, F., Marrinan, J. A. & 11 other authors(1994). The Saccharomyces cerevisiae FKS1 (ETG1) gene encodes an integral membrane protein which is a subunit of 1,3-β-d-glucan synthase. Proc Natl Acad Sci U S A 91, 12907–12911.[CrossRef] [Google Scholar]
  6. Drgonova, J., Drgon, T., Tanaka, K., Kollar, R. Chen G.-C., Ford, R. A., Chan, C. S. M., Takai, Y. & Cabib, E.(1996). Rho1p, a yeast protein at the interface between cell polarization and morphogenesis. Science 272, 277–279.[CrossRef] [Google Scholar]
  7. Errede, B., Cade, R. M., Yashar, B. M., Kamada, Y., Levin, D. E., Irie, K. & Matsomoto, K.(1995). Dynamics and organization of MAP kinase signal pathways. Mol Repr Dev 42, 477–485.[CrossRef] [Google Scholar]
  8. Gray, J. V., Ogas, J. P., Kamada, Y., Stone, M., Levin, D. E. & Herskowitz, I.(1997). A role for the Pkc1 MAP kinase pathway of Saccharomyces cerevisiae in bud emergence and identification of a putative upstream regulator. EMBO J 16, 4924–4937.[CrossRef] [Google Scholar]
  9. Ho, S.-N., Hunt, H. D., Horton, R. M., Pullen, J. K. & Pease, L. R.(1989). Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77, 51–59.[CrossRef] [Google Scholar]
  10. Inoue, S. B., Takewaki, N., Takasuka, T. & 7 other authors(1995). Characterization and gene cloning of 1,3-β-d-glucan synthase from Saccharomyces cerevisiae. Eur J Biochem 231, 845–854.[CrossRef] [Google Scholar]
  11. Irie, K., Takase, M., Lee, K. S., Levin, D. E., Araki, H., Matsumoto, K. & Oshima, Y.(1993).MKK1 and MKK2, which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs, function in the pathway mediated by protein kinase C. Mol Cell Biol 13, 3076–3083. [Google Scholar]
  12. Ito, H., Fukuda, Y., Murata, K. & Kimura, A.(1983). Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153, 163–168. [Google Scholar]
  13. Jacoby, J. J., Nilius, S. M. & Heinisch, J. J.(1998). A screen for upstream components of the yeast protein kinase C signal transduction pathway identifies the product of the SLG1 gene. Mol Gen Genet 258, 148–155.[CrossRef] [Google Scholar]
  14. Jung, U. S. & Levin, D. E.(1999). Genome-wide analysis of gene expression regulated by the yeast cell wall integrity signaling pathway. Mol Microbiol 34, 1049–1057.[CrossRef] [Google Scholar]
  15. Jung, U. S., Sobering, A. K., Romeo, M. J. & Levin, D. E.(2002). Regulation of the yeast Rlm1 transcription factor by the Mpk1 cell wall MAP kinase. Mol Microbiol 46, 781–789.[CrossRef] [Google Scholar]
  16. Kamada, Y., Jung, U. S., Piotrowski, J. & Levin, D. E.(1995). The protein kinase C-activated MAP kinase pathway of Saccharomyces cerevisiae mediates a novel aspect of the heat shock response. Genes Dev 9, 1559–1571.[CrossRef] [Google Scholar]
  17. Kamada, Y., Qadota, H., Python, C. P., Anraku, Y., Ohya, Y. & Levin, D. E.(1996). Activation of yeast protein kinase C by Rho1 GTPase. J Biol Chem 271, 9193–9195.[CrossRef] [Google Scholar]
  18. Ketela, T., Green, R. & Bussey, H.(1999).Saccharomyces cerevisiae Mid2p is a potential cell wall stress sensor and upstream activator of the PKC1-MPK1 cell integrity pathway. J Bacteriol 181, 3330–3340. [Google Scholar]
  19. Kim, Y. J., Francisco, L., Chen, G. C., Marcotte, E. & Chan, C. S. M.(1994). Control of cellular morphogenesis by the Ipl2/Bem2 GTP-activating protein: possible role of protein phosphorylation. J Cell Biol 127, 1381–1394.[CrossRef] [Google Scholar]
  20. Klis, F. M.(1994). Review: cell wall assembly in yeast. Yeast 10, 851–869.[CrossRef] [Google Scholar]
  21. Lee, K. S. & Levin, D. E.(1992). Dominant mutations in a gene encoding a putative protein kinase (BCK1) bypass the requirement for a Saccharomyces cerevisiae protein kinase C homolog. Mol Cell Biol 12, 172–182. [Google Scholar]
  22. Lee, K. S., Irie, K., Gotoh, Y., Watanabe, Y., Araki, H., Nishida, E., Matsumoto, K. & Levin, D. E.(1993). A yeast mitogen-activated protein kinase homolog (Mpk1) mediates signaling by protein kinase C. Mol Cell Biol 13, 3067–3075. [Google Scholar]
  23. Levin, D. E. & Bartlett-Heubusch, E.(1992). Mutants in the S. cerevisiae PKC1 gene display a cell cycle-specific osmotic stability defect. J Cell Biol 116, 1221–1229.[CrossRef] [Google Scholar]
  24. Levin, D. E., Fields, F. O., Kunisawa, R., Bishop, J. M. & Thorner, J.(1990). A candidate protein kinase C gene PKC1, is required for the S. cerevisiae cell cycle. Cell 62, 213–224.[CrossRef] [Google Scholar]
  25. Levin, D. E., Bowers, B., Chen, C., Kamada, Y. & Watanabe, M.(1994). Dissecting the protein kinase C/MAP kinase signaling pathway of Saccharomyces cerevisiae. Cell Mol Biol Res 40, 229–239. [Google Scholar]
  26. Lodder, A. L., Lee, T. K. & Ballester, R.(1999). Characterization of the Wsc1 protein, a putative receptor in the stress response of Saccharomyces cerevisiae. Genetics 152, 1487–1499. [Google Scholar]
  27. Martin, H., Rodriguez-Pachon, J. M., Ruiz, C., Nombela, C. & Molina, M.(2000). Regulatory mechanisms for modulation of signaling through the cell integrity Slt2-mediated pathway in Saccharomyces cerevisiae. J Biol Chem 275, 1511–1519.[CrossRef] [Google Scholar]
  28. Mazur, P., Morin, N., Baginsky, W., El-Sherbeini, M., Clemas, J. A., Nielsen, J. B. & Foor, F.(1995). Differential expression and function of two homologous subunits of yeast 1,3-β-d-glucan synthase. Mol Cell Biol 15, 5671–5681. [Google Scholar]
  29. Nonaka, H., Tanaka, K., Hirano, H., Fujiwara, T., Kohno, H., Umikawa, M., Mino, A. & Takai, Y.(1995). A downstream target of RHO1 small GTP-binding protein is PKC1, a homolog of protein kinase C, which leads to activation of the MAP kinase cascade in Saccharomyces cerevisiae. EMBO J 14, 5931–5938. [Google Scholar]
  30. Ozaki, K., Tanaka, K., Imamura, H., Hihara, T., Kamayema, T., Nonaka, H., Hirano, H., Matsuura, Y. & Takai, Y.(1996). Rom1p and Rom2p are small GDP/GTP exchange proteins (GEPs) for the Rho1p small GTP-binding protein in Saccharomyces cerevisiae. EMBO J 15, 2196–2207. [Google Scholar]
  31. Paravicini, G., Cooper, M., Friedli, L., Smith, D. J., Carpentier, J.-L., Klig, L. S. & Payton, M. A.(1992). The osmotic integrity of the yeast cell requires a functional PKC1 gene product. Mol Cell Biol 12, 4896–4905. [Google Scholar]
  32. Peterson, J., Zheng, Y., Bender, L., Myers, A., Cerione, R. & Bender, A.(1994). Interactions between the bud emergence proteins Bem1 and Bem2 and the Rho-type GTPases in yeast. J Cell Biol 127, 1395–1406.[CrossRef] [Google Scholar]
  33. Philip, B. & Levin, D. E.(2001). Wsc1 and Mid2 are cell surface sensors for cell wall integrity signaling that act through Rom2, a guanine nucleotide exchange factor for Rho1. Mol Cell Biol 21, 271–280.[CrossRef] [Google Scholar]
  34. Qadota, H., Python, C. P., Inoue, S. B., Arisawa, M., Anraku, Y., Zheng, Y., Watanabe, T., Levin, D. E. & Ohya, Y.(1996). Identification of yeast Rho1p GTPase as a regulatory subunit of 1,3-β-glucan synthase. Science 272, 279–281.[CrossRef] [Google Scholar]
  35. Rajavel, M., Philip, B., Buehrer, B. M., Errede, B. & Levin, D. E.(1999). Mid2 is a putative sensor for cell integrity signaling in Saccharomyces cerevisiae. Mol Cell Biol 19, 3969–3976. [Google Scholar]
  36. Ram, A. F. J., Brekelmans, S. S. C., Oehlen, L. J. W. M. & Klis, F. M.(1995). Identification of two cell cycle regulated genes affecting the β-1,3-glucan content of cell wall in Saccharomyces cerevisiae. FEBS Lett 358, 165–170.[CrossRef] [Google Scholar]
  37. Rose, M. D., Winston, F. & Hieter, P.(1990).Methods in Yeast Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  38. Schmidt, A., Bickle, M., Beck, T. & Hall, M.(1997). The yeast phosphatidylinositol kinase homolog TOR2 activates RHO1 and RHO2 via the exchange factor ROM2. Cell 88, 531–542.[CrossRef] [Google Scholar]
  39. Sikorski, R. S. & Hieter, P.(1989). A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122, 19–27. [Google Scholar]
  40. Torres, L., Martin, H., Garcia-Saez, M. I., Arroyo, J., Molina, M., Sanchez, M. & Nombela, C.(1991). A protein kinase gene complements the lytic phenotype of Saccharomyces cerevisiae lyt2 mutants. Mol Microbiol 5, 2845–2854.[CrossRef] [Google Scholar]
  41. Verna, J., Lodder, A., Lee, K., Vagts, A. & Ballester, R.(1997). A family of genes required for the maintenance of cell wall integrity and for the stress response in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 94, 13804–13809.[CrossRef] [Google Scholar]
  42. Watanabe, Y., Takaesu, G., Hagiwara, M., Irie, K. & Matsumoto, K.(1997). Characterization of a serum response factor-like protein in Saccharomyces cerevisiae, Rlm1, which has transcriptional activity regulated by the Mpk1 (Slt2) mitogen-activated protein kinase pathway. Mol Cell Biol 17, 2615–2623. [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.27264-0
Loading
/content/journal/micro/10.1099/mic.0.27264-0
Loading

Data & Media loading...

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