Abstract
In Aspergillus nidulans, germinating conidia undergo multiple rounds of nuclear division before forming a septum. Previous genetic results suggest that the ability to separate nuclear division and septum formation depends upon a threshold level of activity of the cyclin-dependent kinase NIMX(cdk1). Mutations in nimX and nimT, the gene encoding the NIMX(cdk1)-activating phosphatase, have revealed that Tyr-15 phosphorylation is important for determining the timing of the formation of the first septum. Here, we describe a screen for suppressors of nimT23 (snt), designed to identify additional components of the pathway regulating septum formation. We show that a subset of the snt mutants are defective in the temporal regulation of septum formation and in cell cycle checkpoint responses. Molecular characterization of sntA shows that it is allelic to the previously described ankA gene, which encodes the NIMX(cdk1) Tyr-15 kinase. Additional experiments described in this study show that nutritional conditions modulate the timing of septum formation and alter the phenotypes displayed by the snt mutants. A model that suggests that the timing of septum formation is influenced by DNA damage and glucose availability via the sntA and sntB gene products is proposed.
Full Text
The Full Text of this article is available as a PDF (242.6 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Booher R. N., Deshaies R. J., Kirschner M. W. Properties of Saccharomyces cerevisiae wee1 and its differential regulation of p34CDC28 in response to G1 and G2 cyclins. EMBO J. 1993 Sep;12(9):3417–3426. doi: 10.1002/j.1460-2075.1993.tb06016.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cerutti L., Simanis V. Controlling the end of the cell cycle. Curr Opin Genet Dev. 2000 Feb;10(1):65–69. doi: 10.1016/s0959-437x(99)00044-1. [DOI] [PubMed] [Google Scholar]
- De Souza C. P., Ye X. S., Osmani S. A. Checkpoint defects leading to premature mitosis also cause endoreplication of DNA in Aspergillus nidulans. Mol Biol Cell. 1999 Nov;10(11):3661–3674. doi: 10.1091/mbc.10.11.3661. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fantes P. A. Isolation of cell size mutants of a fission yeast by a new selective method: characterization of mutants and implications for division control mechanisms. J Bacteriol. 1981 May;146(2):746–754. doi: 10.1128/jb.146.2.746-754.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gould K. L., Nurse P. Tyrosine phosphorylation of the fission yeast cdc2+ protein kinase regulates entry into mitosis. Nature. 1989 Nov 2;342(6245):39–45. doi: 10.1038/342039a0. [DOI] [PubMed] [Google Scholar]
- Harris S. D., Hamer J. E. sepB: an Aspergillus nidulans gene involved in chromosome segregation and the initiation of cytokinesis. EMBO J. 1995 Nov 1;14(21):5244–5257. doi: 10.1002/j.1460-2075.1995.tb00209.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harris S. D., Hamer L., Sharpless K. E., Hamer J. E. The Aspergillus nidulans sepA gene encodes an FH1/2 protein involved in cytokinesis and the maintenance of cellular polarity. EMBO J. 1997 Jun 16;16(12):3474–3483. doi: 10.1093/emboj/16.12.3474. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harris S. D., Kraus P. R. Regulation of septum formation in Aspergillus nidulans by a DNA damage checkpoint pathway. Genetics. 1998 Mar;148(3):1055–1067. doi: 10.1093/genetics/148.3.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harris S. D., Morrell J. L., Hamer J. E. Identification and characterization of Aspergillus nidulans mutants defective in cytokinesis. Genetics. 1994 Feb;136(2):517–532. doi: 10.1093/genetics/136.2.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harris S. D. The duplication cycle in Aspergillus nidulans. Fungal Genet Biol. 1997 Aug;22(1):1–12. doi: 10.1006/fgbi.1997.0990. [DOI] [PubMed] [Google Scholar]
- Hofmann A. F., Harris S. D. The Aspergillus nidulans uvsB gene encodes an ATM-related kinase required for multiple facets of the DNA damage response. Genetics. 2000 Apr;154(4):1577–1586. doi: 10.1093/genetics/154.4.1577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Humphrey T., Enoch T. Sum1, a highly conserved WD-repeat protein, suppresses S-M checkpoint mutants and inhibits the osmotic stress cell cycle response in fission yeast. Genetics. 1998 Apr;148(4):1731–1742. doi: 10.1093/genetics/148.4.1731. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Käfer E. Meiotic and mitotic recombination in Aspergillus and its chromosomal aberrations. Adv Genet. 1977;19:33–131. doi: 10.1016/s0065-2660(08)60245-x. [DOI] [PubMed] [Google Scholar]
- Lee M. S., Enoch T., Piwnica-Worms H. mik1+ encodes a tyrosine kinase that phosphorylates p34cdc2 on tyrosine 15. J Biol Chem. 1994 Dec 2;269(48):30530–30537. [PubMed] [Google Scholar]
- Lew D. J. Cell-cycle checkpoints that ensure coordination between nuclear and cytoplasmic events in Saccharomyces cerevisiae. Curr Opin Genet Dev. 2000 Feb;10(1):47–53. doi: 10.1016/s0959-437x(99)00051-9. [DOI] [PubMed] [Google Scholar]
- Ma X. J., Lu Q., Grunstein M. A search for proteins that interact genetically with histone H3 and H4 amino termini uncovers novel regulators of the Swe1 kinase in Saccharomyces cerevisiae. Genes Dev. 1996 Jun 1;10(11):1327–1340. doi: 10.1101/gad.10.11.1327. [DOI] [PubMed] [Google Scholar]
- McGowan C. H., Russell P. Human Wee1 kinase inhibits cell division by phosphorylating p34cdc2 exclusively on Tyr15. EMBO J. 1993 Jan;12(1):75–85. doi: 10.1002/j.1460-2075.1993.tb05633.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Millar J. B., McGowan C. H., Lenaers G., Jones R., Russell P. p80cdc25 mitotic inducer is the tyrosine phosphatase that activates p34cdc2 kinase in fission yeast. EMBO J. 1991 Dec;10(13):4301–4309. doi: 10.1002/j.1460-2075.1991.tb05008.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moreno S., Hayles J., Nurse P. Regulation of p34cdc2 protein kinase during mitosis. Cell. 1989 Jul 28;58(2):361–372. doi: 10.1016/0092-8674(89)90850-7. [DOI] [PubMed] [Google Scholar]
- Neufeld T. P., Edgar B. A. Connections between growth and the cell cycle. Curr Opin Cell Biol. 1998 Dec;10(6):784–790. doi: 10.1016/s0955-0674(98)80122-1. [DOI] [PubMed] [Google Scholar]
- O'Connell M. J., Osmani A. H., Morris N. R., Osmani S. A. An extra copy of nimEcyclinB elevates pre-MPF levels and partially suppresses mutation of nimTcdc25 in Aspergillus nidulans. EMBO J. 1992 Jun;11(6):2139–2149. doi: 10.1002/j.1460-2075.1992.tb05273.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osherov N., May G. Conidial germination in Aspergillus nidulans requires RAS signaling and protein synthesis. Genetics. 2000 Jun;155(2):647–656. doi: 10.1093/genetics/155.2.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osmani A. H., McGuire S. L., Osmani S. A. Parallel activation of the NIMA and p34cdc2 cell cycle-regulated protein kinases is required to initiate mitosis in A. nidulans. Cell. 1991 Oct 18;67(2):283–291. doi: 10.1016/0092-8674(91)90180-7. [DOI] [PubMed] [Google Scholar]
- Osmani S. A., May G. S., Morris N. R. Regulation of the mRNA levels of nimA, a gene required for the G2-M transition in Aspergillus nidulans. J Cell Biol. 1987 Jun;104(6):1495–1504. doi: 10.1083/jcb.104.6.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rhind N., Furnari B., Russell P. Cdc2 tyrosine phosphorylation is required for the DNA damage checkpoint in fission yeast. Genes Dev. 1997 Feb 15;11(4):504–511. doi: 10.1101/gad.11.4.504. [DOI] [PubMed] [Google Scholar]
- Russell P., Nurse P. Negative regulation of mitosis by wee1+, a gene encoding a protein kinase homolog. Cell. 1987 May 22;49(4):559–567. doi: 10.1016/0092-8674(87)90458-2. [DOI] [PubMed] [Google Scholar]
- Russell P., Nurse P. cdc25+ functions as an inducer in the mitotic control of fission yeast. Cell. 1986 Apr 11;45(1):145–153. doi: 10.1016/0092-8674(86)90546-5. [DOI] [PubMed] [Google Scholar]
- Sia R. A., Bardes E. S., Lew D. J. Control of Swe1p degradation by the morphogenesis checkpoint. EMBO J. 1998 Nov 16;17(22):6678–6688. doi: 10.1093/emboj/17.22.6678. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Waring R. B., May G. S., Morris N. R. Characterization of an inducible expression system in Aspergillus nidulans using alcA and tubulin-coding genes. Gene. 1989 Jun 30;79(1):119–130. doi: 10.1016/0378-1119(89)90097-8. [DOI] [PubMed] [Google Scholar]
- Wolkow T. D., Harris S. D., Hamer J. E. Cytokinesis in Aspergillus nidulans is controlled by cell size, nuclear positioning and mitosis. J Cell Sci. 1996 Aug;109(Pt 8):2179–2188. doi: 10.1242/jcs.109.8.2179. [DOI] [PubMed] [Google Scholar]
- Wu L., Russell P. Nim1 kinase promotes mitosis by inactivating Wee1 tyrosine kinase. Nature. 1993 Jun 24;363(6431):738–741. doi: 10.1038/363738a0. [DOI] [PubMed] [Google Scholar]
- Ye X. S., Fincher R. R., Tang A., Osmani S. A. The G2/M DNA damage checkpoint inhibits mitosis through Tyr15 phosphorylation of p34cdc2 in Aspergillus nidulans. EMBO J. 1997 Jan 2;16(1):182–192. doi: 10.1093/emboj/16.1.182. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zeng Y., Forbes K. C., Wu Z., Moreno S., Piwnica-Worms H., Enoch T. Replication checkpoint requires phosphorylation of the phosphatase Cdc25 by Cds1 or Chk1. Nature. 1998 Oct 1;395(6701):507–510. doi: 10.1038/26766. [DOI] [PubMed] [Google Scholar]