Abstract
Neurospora crassa is a filamentous fungus that grows on semisolid media by forming spreading colonies. Mutations at several loci prevent this spreading growth. cot-1 is a temperature sensitive mutant of N.crassa that exhibits restricted colonial growth. At temperatures above 32 degrees C colonies are compact while at lower temperatures growth is indistinguishable from that of the wild type. Restricted colonial growth is due to a defect in hyphal tip elongation and a concomitant increase in hyphal branching. We have isolated a genomic cosmid clone containing the wild type allele of cot-1 by complementation. Sequence analyses suggested that cot-1 encodes a member of the cAMP-dependent protein kinase family. Strains in which we disrupted cot-1 are viable but display restricted colonial growth. Duplication, by ectopic integration of a promoter-containing fragment which includes the first one-third (209 codons) of the structural gene, unexpectedly resulted in restricted colonial growth. Our results suggest that an active COT1 kinase is required for one or more events essential for hyphal elongation.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bahman M., Buck V., White A., Rosamond J. Characterisation of the CDC7 gene product of Saccharomyces cerevisiae as a protein kinase needed for the initiation of mitotic DNA synthesis. Biochim Biophys Acta. 1988 Dec 20;951(2-3):335–343. doi: 10.1016/0167-4781(88)90104-2. [DOI] [PubMed] [Google Scholar]
- Bellman R., Lehman S. ON THE CONTINUOUS GOLD-MINING EQUATION. Proc Natl Acad Sci U S A. 1954 Feb;40(2):115–119. doi: 10.1073/pnas.40.2.115. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
- Berlin V., Yanofsky C. Protein changes during the asexual cycle of Neurospora crassa. Mol Cell Biol. 1985 Apr;5(4):839–848. doi: 10.1128/mcb.5.4.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cullen D., Leong S. A., Wilson L. J., Henner D. J. Transformation of Aspergillus nidulans with the hygromycin-resistance gene, hph. Gene. 1987;57(1):21–26. doi: 10.1016/0378-1119(87)90172-7. [DOI] [PubMed] [Google Scholar]
- Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fincham J. R. Transformation in fungi. Microbiol Rev. 1989 Mar;53(1):148–170. doi: 10.1128/mr.53.1.148-170.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grove S. N., Bracker C. E. Protoplasmic organization of hyphal tips among fungi: vesicles and Spitzenkörper. J Bacteriol. 1970 Nov;104(2):989–1009. doi: 10.1128/jb.104.2.989-1009.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hager K. M., Yanofsky C. Genes expressed during conidiation in Neurospora crassa: molecular characterization of con-13. Gene. 1990 Dec 15;96(2):153–159. doi: 10.1016/0378-1119(90)90247-o. [DOI] [PubMed] [Google Scholar]
- Hanks S. K., Quinn A. M., Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988 Jul 1;241(4861):42–52. doi: 10.1126/science.3291115. [DOI] [PubMed] [Google Scholar]
- Johnston L. H., Eberly S. L., Chapman J. W., Araki H., Sugino A. The product of the Saccharomyces cerevisiae cell cycle gene DBF2 has homology with protein kinases and is periodically expressed in the cell cycle. Mol Cell Biol. 1990 Apr;10(4):1358–1366. doi: 10.1128/mcb.10.4.1358. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knighton D. R., Zheng J. H., Ten Eyck L. F., Ashford V. A., Xuong N. H., Taylor S. S., Sowadski J. M. Crystal structure of the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. Science. 1991 Jul 26;253(5018):407–414. doi: 10.1126/science.1862342. [DOI] [PubMed] [Google Scholar]
- Knighton D. R., Zheng J. H., Ten Eyck L. F., Xuong N. H., Taylor S. S., Sowadski J. M. Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. Science. 1991 Jul 26;253(5018):414–420. doi: 10.1126/science.1862343. [DOI] [PubMed] [Google Scholar]
- Metzenberg R. L., Stevens J. N., Selker E. U., Morzycka-Wroblewska E. Identification and chromosomal distribution of 5S rRNA genes in Neurospora crassa. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2067–2071. doi: 10.1073/pnas.82.7.2067. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Orbach M. J., Porro E. B., Yanofsky C. Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker. Mol Cell Biol. 1986 Jul;6(7):2452–2461. doi: 10.1128/mcb.6.7.2452. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paluh J. L., Orbach M. J., Legerton T. L., Yanofsky C. The cross-pathway control gene of Neurospora crassa, cpc-1, encodes a protein similar to GCN4 of yeast and the DNA-binding domain of the oncogene v-jun-encoded protein. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3728–3732. doi: 10.1073/pnas.85.11.3728. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Patterson M., Sclafani R. A., Fangman W. L., Rosamond J. Molecular characterization of cell cycle gene CDC7 from Saccharomyces cerevisiae. Mol Cell Biol. 1986 May;6(5):1590–1598. doi: 10.1128/mcb.6.5.1590. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perkins D. D., Newmeyer D., Taylor C. W., Bennett D. C. New markers and map sequences in Neurospora crassa, with a description of mapping by duplication coverage, and of multiple translocation stocks for testing linkage. Genetica. 1969;40(3):247–278. doi: 10.1007/BF01787357. [DOI] [PubMed] [Google Scholar]
- Perkins D. D., Radford A., Newmeyer D., Björkman M. Chromosomal loci of Neurospora crassa. Microbiol Rev. 1982 Dec;46(4):426–570. doi: 10.1128/mr.46.4.426-570.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rambosek J., Leach J. Recombinant DNA in filamentous fungi: progress and prospects. Crit Rev Biotechnol. 1987;6(4):357–393. doi: 10.3109/07388558709089387. [DOI] [PubMed] [Google Scholar]
- Sachs M. S., Yanofsky C. Developmental expression of genes involved in conidiation and amino acid biosynthesis in Neurospora crassa. Dev Biol. 1991 Nov;148(1):117–128. doi: 10.1016/0012-1606(91)90322-t. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmit J. C., Brody S. Biochemical genetics of Neurospora crassa conidial germination. Bacteriol Rev. 1976 Mar;40(1):1–41. doi: 10.1128/br.40.1.1-41.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Subbiah S., Harrison S. C. A method for multiple sequence alignment with gaps. J Mol Biol. 1989 Oct 20;209(4):539–548. doi: 10.1016/0022-2836(89)90592-5. [DOI] [PubMed] [Google Scholar]
- Tatum E. L., Barratt R. W., Cutter V. M., Jr Chemical Induction of Colonial Paramorphs in Neurospora and Syncephalastrum. Science. 1949 May 20;109(2838):509–511. doi: 10.1126/science.109.2838.509. [DOI] [PubMed] [Google Scholar]
- Taylor S. S. cAMP-dependent protein kinase. Model for an enzyme family. J Biol Chem. 1989 May 25;264(15):8443–8446. [PubMed] [Google Scholar]
- Toyn J. H., Araki H., Sugino A., Johnston L. H. The cell-cycle-regulated budding yeast gene DBF2, encoding a putative protein kinase, has a homologue that is not under cell-cycle control. Gene. 1991 Jul 31;104(1):63–70. doi: 10.1016/0378-1119(91)90465-n. [DOI] [PubMed] [Google Scholar]
- Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
- Vollmer S. J., Yanofsky C. Efficient cloning of genes of Neurospora crassa. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4869–4873. doi: 10.1073/pnas.83.13.4869. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yarden O., Yanofsky C. Chitin synthase 1 plays a major role in cell wall biogenesis in Neurospora crassa. Genes Dev. 1991 Dec;5(12B):2420–2430. doi: 10.1101/gad.5.12b.2420. [DOI] [PubMed] [Google Scholar]