Skip to main content
PMC home page
Genetics logoLink to Genetics
. 1998 Jul;149(3):1303–1321. doi: 10.1093/genetics/149.3.1303

A genetic screen for temperature-sensitive cell-division mutants of Caenorhabditis elegans.

K F O'Connell 1, C M Leys 1, J G White 1
PMCID: PMC1460235  PMID: 9649522

Abstract

A novel screen to isolate conditional cell-division mutants in Caenorhabditis elegans has been developed. The screen is based on the phenotypes associated with existing cell-division mutations: some disrupt postembryonic divisions and affect formation of the gonad and ventral nerve cord-resulting in sterile, uncoordinated animals-while others affect embryonic divisions and result in lethality. We obtained 19 conditional mutants that displayed these phenotypes when shifted to the restrictive temperature at the appropriate developmental stage. Eighteen of these mutations have been mapped; 17 proved to be single alleles of newly identified genes, while 1 proved to be an allele of a previously identified gene. Genetic tests on the embryonic lethal phenotypes indicated that for 13 genes, embryogenesis required maternal expression, while for 6, zygotic expression could suffice. In all cases, maternal expression of wild-type activity was found to be largely sufficient for embryogenesis. Cytological analysis revealed that 10 mutants possessed embryonic cell-division defects, including failure to properly segregate DNA, failure to assemble a mitotic spindle, late cytokinesis defects, prolonged cell cycles, and improperly oriented mitotic spindles. We conclude that this approach can be used to identify mutations that affect various aspects of the cell-division cycle.

Full Text

The Full Text of this article is available as a PDF (406.5 KB).

Selected References

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

  1. Aroian R. V., Field C., Pruliere G., Kenyon C., Alberts B. M. Isolation of actin-associated proteins from Caenorhabditis elegans oocytes and their localization in the early embryo. EMBO J. 1997 Apr 1;16(7):1541–1549. doi: 10.1093/emboj/16.7.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Assaad F. F., Mayer U., Wanner G., Jürgens G. The KEULE gene is involved in cytokinesis in Arabidopsis. Mol Gen Genet. 1996 Dec 13;253(3):267–277. doi: 10.1007/pl00008594. [DOI] [PubMed] [Google Scholar]
  3. Bhat M. A., Philp A. V., Glover D. M., Bellen H. J. Chromatid segregation at anaphase requires the barren product, a novel chromosome-associated protein that interacts with Topoisomerase II. Cell. 1996 Dec 13;87(6):1103–1114. doi: 10.1016/s0092-8674(00)81804-8. [DOI] [PubMed] [Google Scholar]
  4. Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cassada R., Isnenghi E., Culotti M., von Ehrenstein G. Genetic analysis of temperature-sensitive embryogenesis mutants in Caenorhabditis elegans. Dev Biol. 1981 May;84(1):193–205. doi: 10.1016/0012-1606(81)90383-3. [DOI] [PubMed] [Google Scholar]
  6. Cheng N. N., Kirby C. M., Kemphues K. J. Control of cleavage spindle orientation in Caenorhabditis elegans: the role of the genes par-2 and par-3. Genetics. 1995 Feb;139(2):549–559. doi: 10.1093/genetics/139.2.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Conrad G. W., Williams D. C., Turner F. R., Newrock K. M., Raff R. A. Microfilaments in the polar lobe constriction of fertilized eggs of Ilyanassa obsoleta. J Cell Biol. 1973 Oct;59(1):228–233. doi: 10.1083/jcb.59.1.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Eeckman F. H., Durbin R. ACeDB and macace. Methods Cell Biol. 1995;48:583–605. [PubMed] [Google Scholar]
  9. Etemad-Moghadam B., Guo S., Kemphues K. J. Asymmetrically distributed PAR-3 protein contributes to cell polarity and spindle alignment in early C. elegans embryos. Cell. 1995 Dec 1;83(5):743–752. doi: 10.1016/0092-8674(95)90187-6. [DOI] [PubMed] [Google Scholar]
  10. Gatti M., Baker B. S. Genes controlling essential cell-cycle functions in Drosophila melanogaster. Genes Dev. 1989 Apr;3(4):438–453. doi: 10.1101/gad.3.4.438. [DOI] [PubMed] [Google Scholar]
  11. Hird S. N., White J. G. Cortical and cytoplasmic flow polarity in early embryonic cells of Caenorhabditis elegans. J Cell Biol. 1993 Jun;121(6):1343–1355. doi: 10.1083/jcb.121.6.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hirsh D., Vanderslice R. Temperature-sensitive developmental mutants of Caenorhabditis elegans. Dev Biol. 1976 Mar;49(1):220–235. doi: 10.1016/0012-1606(76)90268-2. [DOI] [PubMed] [Google Scholar]
  13. Hodgkin J., Horvitz H. R., Brenner S. Nondisjunction Mutants of the Nematode CAENORHABDITIS ELEGANS. Genetics. 1979 Jan;91(1):67–94. doi: 10.1093/genetics/91.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Horvitz H. R., Sulston J. E. Isolation and genetic characterization of cell-lineage mutants of the nematode Caenorhabditis elegans. Genetics. 1980 Oct;96(2):435–454. doi: 10.1093/genetics/96.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hyman A. A., White J. G. Determination of cell division axes in the early embryogenesis of Caenorhabditis elegans. J Cell Biol. 1987 Nov;105(5):2123–2135. doi: 10.1083/jcb.105.5.2123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kemphues K. J., Kusch M., Wolf N. Maternal-effect lethal mutations on linkage group II of Caenorhabditis elegans. Genetics. 1988 Dec;120(4):977–986. doi: 10.1093/genetics/120.4.977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kemphues K. J., Wolf N., Wood W. B., Hirsh D. Two loci required for cytoplasmic organization in early embryos of Caenorhabditis elegans. Dev Biol. 1986 Feb;113(2):449–460. doi: 10.1016/0012-1606(86)90180-6. [DOI] [PubMed] [Google Scholar]
  18. Kipreos E. T., Lander L. E., Wing J. P., He W. W., Hedgecock E. M. cul-1 is required for cell cycle exit in C. elegans and identifies a novel gene family. Cell. 1996 Jun 14;85(6):829–839. doi: 10.1016/s0092-8674(00)81267-2. [DOI] [PubMed] [Google Scholar]
  19. Lewis J. A., Fleming J. T. Basic culture methods. Methods Cell Biol. 1995;48:3–29. [PubMed] [Google Scholar]
  20. McKim K. S., Starr T., Rose A. M. Genetic and molecular analysis of the dpy-14 region in Caenorhabditis elegans. Mol Gen Genet. 1992 May;233(1-2):241–251. doi: 10.1007/BF00587585. [DOI] [PubMed] [Google Scholar]
  21. McKim K. S., Starr T., Rose A. M. Genetic and molecular analysis of the dpy-14 region in Caenorhabditis elegans. Mol Gen Genet. 1992 May;233(1-2):241–251. doi: 10.1007/BF00587585. [DOI] [PubMed] [Google Scholar]
  22. Nasmyth K., Nurse P. Cell division cycle mutants altered in DNA replication and mitosis in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1981;182(1):119–124. doi: 10.1007/BF00422777. [DOI] [PubMed] [Google Scholar]
  23. Nurse P., Thuriaux P., Nasmyth K. Genetic control of the cell division cycle in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1976 Jul 23;146(2):167–178. doi: 10.1007/BF00268085. [DOI] [PubMed] [Google Scholar]
  24. Priess J. R., Schnabel H., Schnabel R. The glp-1 locus and cellular interactions in early C. elegans embryos. Cell. 1987 Nov 20;51(4):601–611. doi: 10.1016/0092-8674(87)90129-2. [DOI] [PubMed] [Google Scholar]
  25. Samejima I., Matsumoto T., Nakaseko Y., Beach D., Yanagida M. Identification of seven new cut genes involved in Schizosaccharomyces pombe mitosis. J Cell Sci. 1993 May;105(Pt 1):135–143. doi: 10.1242/jcs.105.1.135. [DOI] [PubMed] [Google Scholar]
  26. Seydoux G., Fire A. Soma-germline asymmetry in the distributions of embryonic RNAs in Caenorhabditis elegans. Development. 1994 Oct;120(10):2823–2834. doi: 10.1242/dev.120.10.2823. [DOI] [PubMed] [Google Scholar]
  27. Strome S., Wood W. B. Generation of asymmetry and segregation of germ-line granules in early C. elegans embryos. Cell. 1983 Nov;35(1):15–25. doi: 10.1016/0092-8674(83)90203-9. [DOI] [PubMed] [Google Scholar]
  28. Sulston J. E., Horvitz H. R. Abnormal cell lineages in mutants of the nematode Caenorhabditis elegans. Dev Biol. 1981 Feb;82(1):41–55. doi: 10.1016/0012-1606(81)90427-9. [DOI] [PubMed] [Google Scholar]
  29. Sulston J. E., Horvitz H. R. Post-embryonic cell lineages of the nematode, Caenorhabditis elegans. Dev Biol. 1977 Mar;56(1):110–156. doi: 10.1016/0012-1606(77)90158-0. [DOI] [PubMed] [Google Scholar]
  30. Thomas C., DeVries P., Hardin J., White J. Four-dimensional imaging: computer visualization of 3D movements in living specimens. Science. 1996 Aug 2;273(5275):603–607. doi: 10.1126/science.273.5275.603. [DOI] [PubMed] [Google Scholar]
  31. Uemura T., Ohkura H., Adachi Y., Morino K., Shiozaki K., Yanagida M. DNA topoisomerase II is required for condensation and separation of mitotic chromosomes in S. pombe. Cell. 1987 Sep 11;50(6):917–925. doi: 10.1016/0092-8674(87)90518-6. [DOI] [PubMed] [Google Scholar]
  32. Waddle J. A., Cooper J. A., Waterston R. H. Transient localized accumulation of actin in Caenorhabditis elegans blastomeres with oriented asymmetric divisions. Development. 1994 Aug;120(8):2317–2328. doi: 10.1242/dev.120.8.2317. [DOI] [PubMed] [Google Scholar]
  33. Ward S., Miwa J. Characterization of temperature-sensitive, fertilization-defective mutants of the nematode caenorhabditis elegans. Genetics. 1978 Feb;88(2):285–303. doi: 10.1093/genetics/88.2.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wheatley S. P., Wang Y. Midzone microtubule bundles are continuously required for cytokinesis in cultured epithelial cells. J Cell Biol. 1996 Nov;135(4):981–989. doi: 10.1083/jcb.135.4.981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Williams B. C., Riedy M. F., Williams E. V., Gatti M., Goldberg M. L. The Drosophila kinesin-like protein KLP3A is a midbody component required for central spindle assembly and initiation of cytokinesis. J Cell Biol. 1995 May;129(3):709–723. doi: 10.1083/jcb.129.3.709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wood W. B., Hecht R., Carr S., Vanderslice R., Wolf N., Hirsh D. Parental effects and phenotypic characterization of mutations that affect early development in Caenorhabditis elegans. Dev Biol. 1980 Feb;74(2):446–469. doi: 10.1016/0012-1606(80)90445-5. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES