Skip to main content
Genetics logoLink to Genetics
. 2003 Oct;165(2):517–529. doi: 10.1093/genetics/165.2.517

Yeast Nap1-binding protein Nbp2p is required for mitotic growth at high temperatures and for cell wall integrity.

Kentaro Ohkuni 1, Asuko Okuda 1, Akihiko Kikuchi 1
PMCID: PMC1462787  PMID: 14573466

Abstract

Nbp2p is a Nap1-binding protein in Saccharomyces cerevisiae identified by its interaction with Nap1 by a two-hybrid system. NBP2 encodes a novel protein consisting of 236 amino acids with a Src homology 3 (SH3) domain. We showed that NBP2 functions to promote mitotic cell growth at high temperatures and cell wall integrity. Loss of Nbp2 results in cell death at high temperatures and in sensitivity to calcofluor white. Cell death at high temperature is thought not to be due to a weakened cell wall. Additionally, we have isolated several type-2C serine threonine protein phosphatases (PTCs) as multicopy suppressors and MAP kinase-kinase (MAPKK), related to the yeast PKC MAPK pathway, as deletion suppressors of the nbp2Delta mutant. Screening for deletion suppressors is a new genetic approach to identify and characterize additional proteins in the Nbp2-dependent pathway. Genetic analyses suggested that Ptc1, which interacts with Nbp2 by the two-hybrid system, acts downstream of Nbp2 and that cells lacking the function of Nbp2 prefer to lose Mkk1, but the PKC MAPK pathway itself is indispensable when Nbp2 is deleted at high temperature.

Full Text

The Full Text of this article is available as a PDF (459.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boles E., Schulte F., Miosga T., Freidel K., Schlüter E., Zimmermann F. K., Hollenberg C. P., Heinisch J. J. Characterization of a glucose-repressed pyruvate kinase (Pyk2p) in Saccharomyces cerevisiae that is catalytically insensitive to fructose-1,6-bisphosphate. J Bacteriol. 1997 May;179(9):2987–2993. doi: 10.1128/jb.179.9.2987-2993.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Botstein D., Falco S. C., Stewart S. E., Brennan M., Scherer S., Stinchcomb D. T., Struhl K., Davis R. W. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. doi: 10.1016/0378-1119(79)90004-0. [DOI] [PubMed] [Google Scholar]
  3. Burns N., Grimwade B., Ross-Macdonald P. B., Choi E. Y., Finberg K., Roeder G. S., Snyder M. Large-scale analysis of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae. Genes Dev. 1994 May 1;8(9):1087–1105. doi: 10.1101/gad.8.9.1087. [DOI] [PubMed] [Google Scholar]
  4. Cabib E., Duran A. Simple and sensitive procedure for screening yeast mutants that lyse at nonpermissive temperatures. J Bacteriol. 1975 Dec;124(3):1604–1606. doi: 10.1128/jb.124.3.1604-1606.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang L., Loranger S. S., Mizzen C., Ernst S. G., Allis C. D., Annunziato A. T. Histones in transit: cytosolic histone complexes and diacetylation of H4 during nucleosome assembly in human cells. Biochemistry. 1997 Jan 21;36(3):469–480. doi: 10.1021/bi962069i. [DOI] [PubMed] [Google Scholar]
  6. Chien C. T., Bartel P. L., Sternglanz R., Fields S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9578–9582. doi: 10.1073/pnas.88.21.9578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Evangelista Marie, Pruyne David, Amberg David C., Boone Charles, Bretscher Aanthony. Formins direct Arp2/3-independent actin filament assembly to polarize cell growth in yeast. Nat Cell Biol. 2002 Jan;4(1):32–41. doi: 10.1038/ncb718. [DOI] [PubMed] [Google Scholar]
  8. Fonzi W. A., Irwin M. Y. Isogenic strain construction and gene mapping in Candida albicans. Genetics. 1993 Jul;134(3):717–728. doi: 10.1093/genetics/134.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Irie K., Takase M., Lee K. S., Levin D. E., Araki H., Matsumoto K., Oshima Y. 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. 1993 May;13(5):3076–3083. doi: 10.1128/mcb.13.5.3076. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ishimi Y., Yasuda H., Hirosumi J., Hanaoka F., Yamada M. A protein which facilitates assembly of nucleosome-like structures in vitro in mammalian cells. J Biochem. 1983 Sep;94(3):735–744. doi: 10.1093/oxfordjournals.jbchem.a134414. [DOI] [PubMed] [Google Scholar]
  11. Ito T., Bulger M., Pazin M. J., Kobayashi R., Kadonaga J. T. ACF, an ISWI-containing and ATP-utilizing chromatin assembly and remodeling factor. Cell. 1997 Jul 11;90(1):145–155. doi: 10.1016/s0092-8674(00)80321-9. [DOI] [PubMed] [Google Scholar]
  12. Ito T., Tashiro K., Muta S., Ozawa R., Chiba T., Nishizawa M., Yamamoto K., Kuhara S., Sakaki Y. Toward a protein-protein interaction map of the budding yeast: A comprehensive system to examine two-hybrid interactions in all possible combinations between the yeast proteins. Proc Natl Acad Sci U S A. 2000 Feb 1;97(3):1143–1147. doi: 10.1073/pnas.97.3.1143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jiang B., Ram A. F., Sheraton J., Klis F. M., Bussey H. Regulation of cell wall beta-glucan assembly: PTC1 negatively affects PBS2 action in a pathway that includes modulation of EXG1 transcription. Mol Gen Genet. 1995 Aug 21;248(3):260–269. doi: 10.1007/BF02191592. [DOI] [PubMed] [Google Scholar]
  14. Kaeberlein Matt, Guarente Leonard. Saccharomyces cerevisiae MPT5 and SSD1 function in parallel pathways to promote cell wall integrity. Genetics. 2002 Jan;160(1):83–95. doi: 10.1093/genetics/160.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kawase H., Okuwaki M., Miyaji M., Ohba R., Handa H., Ishimi Y., Fujii-Nakata T., Kikuchi A., Nagata K. NAP-I is a functional homologue of TAF-I that is required for replication and transcription of the adenovirus genome in a chromatin-like structure. Genes Cells. 1996 Dec;1(12):1045–1056. doi: 10.1046/j.1365-2443.1996.d01-223.x. [DOI] [PubMed] [Google Scholar]
  16. Kellogg D. R., Kikuchi A., Fujii-Nakata T., Turck C. W., Murray A. W. Members of the NAP/SET family of proteins interact specifically with B-type cyclins. J Cell Biol. 1995 Aug;130(3):661–673. doi: 10.1083/jcb.130.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kellogg D. R., Murray A. W. NAP1 acts with Clb1 to perform mitotic functions and to suppress polar bud growth in budding yeast. J Cell Biol. 1995 Aug;130(3):675–685. doi: 10.1083/jcb.130.3.675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Levin D. E., Errede B. The proliferation of MAP kinase signaling pathways in yeast. Curr Opin Cell Biol. 1995 Apr;7(2):197–202. doi: 10.1016/0955-0674(95)80028-x. [DOI] [PubMed] [Google Scholar]
  19. Matsumoto K., Nagata K., Miyaji-Yamaguchi M., Kikuchi A., Tsujimoto M. Sperm chromatin decondensation by template activating factor I through direct interaction with basic proteins. Mol Cell Biol. 1999 Oct;19(10):6940–6952. doi: 10.1128/mcb.19.10.6940. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mayer B. J. SH3 domains: complexity in moderation. J Cell Sci. 2001 Apr;114(Pt 7):1253–1263. doi: 10.1242/jcs.114.7.1253. [DOI] [PubMed] [Google Scholar]
  21. Nagata K., Kawase H., Handa H., Yano K., Yamasaki M., Ishimi Y., Okuda A., Kikuchi A., Matsumoto K. Replication factor encoded by a putative oncogene, set, associated with myeloid leukemogenesis. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4279–4283. doi: 10.1073/pnas.92.10.4279. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nagata K., Saito S., Okuwaki M., Kawase H., Furuya A., Kusano A., Hanai N., Okuda A., Kikuchi A. Cellular localization and expression of template-activating factor I in different cell types. Exp Cell Res. 1998 May 1;240(2):274–281. doi: 10.1006/excr.1997.3930. [DOI] [PubMed] [Google Scholar]
  23. Ohkuni K., Yamashita I. A transcriptional autoregulatory loop for KIN28-CCL1 and SRB10-SRB11, each encoding RNA polymerase II CTD kinase-cyclin pair, stimulates the meiotic development of S. cerevisiae. Yeast. 2000 Jun 30;16(9):829–846. doi: 10.1002/1097-0061(20000630)16:9<829::AID-YEA581>3.0.CO;2-6. [DOI] [PubMed] [Google Scholar]
  24. Paravicini G., Cooper M., Friedli L., Smith D. J., Carpentier J. L., Klig L. S., Payton M. A. The osmotic integrity of the yeast cell requires a functional PKC1 gene product. Mol Cell Biol. 1992 Nov;12(11):4896–4905. doi: 10.1128/mcb.12.11.4896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Parent S. A., Fenimore C. M., Bostian K. A. Vector systems for the expression, analysis and cloning of DNA sequences in S. cerevisiae. Yeast. 1985 Dec;1(2):83–138. doi: 10.1002/yea.320010202. [DOI] [PubMed] [Google Scholar]
  26. Poch O. Conservation of a putative inhibitory domain in the GAL4 family members. Gene. 1997 Jan 15;184(2):229–235. doi: 10.1016/s0378-1119(96)00602-6. [DOI] [PubMed] [Google Scholar]
  27. Saito S., Miyaji-Yamaguchi M., Shimoyama T., Nagata K. Functional domains of template-activating factor-I as a protein phosphatase 2A inhibitor. Biochem Biophys Res Commun. 1999 Jun 7;259(2):471–475. doi: 10.1006/bbrc.1999.0790. [DOI] [PubMed] [Google Scholar]
  28. Shimizu Y., Akashi T., Okuda A., Kikuchi A., Fukui K. NBP1 (Nap1 binding protein 1), an essential gene for G2/M transition of Saccharomyces cerevisiae, encodes a protein of distinct sub-nuclear localization. Gene. 2000 Apr 4;246(1-2):395–404. doi: 10.1016/s0378-1119(00)00067-6. [DOI] [PubMed] [Google Scholar]
  29. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sreenivasan A., Kellogg D. The elm1 kinase functions in a mitotic signaling network in budding yeast. Mol Cell Biol. 1999 Dec;19(12):7983–7994. doi: 10.1128/mcb.19.12.7983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sutton A., Immanuel D., Arndt K. T. The SIT4 protein phosphatase functions in late G1 for progression into S phase. Mol Cell Biol. 1991 Apr;11(4):2133–2148. doi: 10.1128/mcb.11.4.2133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tong A. H., Evangelista M., Parsons A. B., Xu H., Bader G. D., Pagé N., Robinson M., Raghibizadeh S., Hogue C. W., Bussey H. Systematic genetic analysis with ordered arrays of yeast deletion mutants. Science. 2001 Dec 14;294(5550):2364–2368. doi: 10.1126/science.1065810. [DOI] [PubMed] [Google Scholar]
  33. Uesono Y., Fujita A., Toh-e A., Kikuchi Y. The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast. Gene. 1994 May 27;143(1):135–138. doi: 10.1016/0378-1119(94)90618-1. [DOI] [PubMed] [Google Scholar]
  34. Uesono Y., Toh-e A., Kikuchi Y. Ssd1p of Saccharomyces cerevisiae associates with RNA. J Biol Chem. 1997 Jun 27;272(26):16103–16109. doi: 10.1074/jbc.272.26.16103. [DOI] [PubMed] [Google Scholar]
  35. Uetz P., Giot L., Cagney G., Mansfield T. A., Judson R. S., Knight J. R., Lockshon D., Narayan V., Srinivasan M., Pochart P. A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. Nature. 2000 Feb 10;403(6770):623–627. doi: 10.1038/35001009. [DOI] [PubMed] [Google Scholar]
  36. Vojtek A. B., Hollenberg S. M., Cooper J. A. Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell. 1993 Jul 16;74(1):205–214. doi: 10.1016/0092-8674(93)90307-c. [DOI] [PubMed] [Google Scholar]
  37. Warmka J., Hanneman J., Lee J., Amin D., Ota I. Ptc1, a type 2C Ser/Thr phosphatase, inactivates the HOG pathway by dephosphorylating the mitogen-activated protein kinase Hog1. Mol Cell Biol. 2001 Jan;21(1):51–60. doi: 10.1128/MCB.21.1.51-60.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yang P., Pimental R., Lai H., Marcus S. Direct activation of the fission yeast PAK Shk1 by the novel SH3 domain protein, Skb5. J Biol Chem. 1999 Dec 17;274(51):36052–36057. doi: 10.1074/jbc.274.51.36052. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES