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. 1993 Feb 2;120(4):959–968. doi: 10.1083/jcb.120.4.959

BIMA, a TPR-containing protein required for mitosis, localizes to the spindle pole body in Aspergillus nidulans

PMCID: PMC2200070  PMID: 8432735

Abstract

The Aspergillus nidulans bimA gene is required for mitosis. Loss of function mutations in bimA cause cells to arrest growth with condensed chromatin and a short, metaphaselike mitotic spindle. bimA is a member of a gene family defined by a repeated motif called the Tetratrico Peptide Repeat (TPR), which is found in genes from bacteria, yeast and insects. Several yeast TPR genes are also required for mitosis, including Saccharomyces cerevisiae CDC27 and Schizosaccharomyces pombe nuc2+, which appear to be functional homologs of bimA. We have developed antisera specific to the bimA protein (BIMA) and have characterized BIMA by western blot and immunocytochemical analyses. BIMA is heterogeneous in apparent molecular weight, consisting of a major 90-kD species and at least two minor species of approximately 105 kD. The results of BIMA localization by immunofluorescence microscopy depend on the level of BIMA expression. Overexpression of BIMA, which had no deleterious affect on growth or mitosis, resulted in localization of BIMA on or throughout most nuclei. Nuclear staining was granular, and overlapped but was not completely coincident with DNA staining by DAPI. In contrast, when expressed at normal levels, BIMA colocalized with the spindle pole body (SPB). BIMA localized to the SPB in a cell cycle independent manner. These results show that BIMA is either associated with or is a component of the SPB, and they suggest that BIMA functions at the spindle poles to promote the onset of anaphase.

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Selected References

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  1. Alfa C. E., Ducommun B., Beach D., Hyams J. S. Distinct nuclear and spindle pole body population of cyclin-cdc2 in fission yeast. Nature. 1990 Oct 18;347(6294):680–682. doi: 10.1038/347680a0. [DOI] [PubMed] [Google Scholar]
  2. Bergen L. G., Morris N. R. Kinetics of the nuclear division cycle of Aspergillus nidulans. J Bacteriol. 1983 Oct;156(1):155–160. doi: 10.1128/jb.156.1.155-160.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dalrymple M. A., Petersen-Bjorn S., Friesen J. D., Beggs J. D. The product of the PRP4 gene of S. cerevisiae shows homology to beta subunits of G proteins. Cell. 1989 Sep 8;58(5):811–812. doi: 10.1016/0092-8674(89)90930-6. [DOI] [PubMed] [Google Scholar]
  4. Dunphy W. G., Kumagai A. The cdc25 protein contains an intrinsic phosphatase activity. Cell. 1991 Oct 4;67(1):189–196. doi: 10.1016/0092-8674(91)90582-j. [DOI] [PubMed] [Google Scholar]
  5. Engle D. B., Doonan J. H., Morris N. R. Cell-cycle modulation of MPM-2-specific spindle pole body phosphorylation in Aspergillus nidulans. Cell Motil Cytoskeleton. 1988;10(3):434–437. doi: 10.1002/cm.970100310. [DOI] [PubMed] [Google Scholar]
  6. Forsburg S. L., Nurse P. Cell cycle regulation in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. Annu Rev Cell Biol. 1991;7:227–256. doi: 10.1146/annurev.cb.07.110191.001303. [DOI] [PubMed] [Google Scholar]
  7. Gautier J., Solomon M. J., Booher R. N., Bazan J. F., Kirschner M. W. cdc25 is a specific tyrosine phosphatase that directly activates p34cdc2. Cell. 1991 Oct 4;67(1):197–211. doi: 10.1016/0092-8674(91)90583-k. [DOI] [PubMed] [Google Scholar]
  8. Goebl M., Yanagida M. The TPR snap helix: a novel protein repeat motif from mitosis to transcription. Trends Biochem Sci. 1991 May;16(5):173–177. doi: 10.1016/0968-0004(91)90070-c. [DOI] [PubMed] [Google Scholar]
  9. Hirano T., Hiraoka Y., Yanagida M. A temperature-sensitive mutation of the Schizosaccharomyces pombe gene nuc2+ that encodes a nuclear scaffold-like protein blocks spindle elongation in mitotic anaphase. J Cell Biol. 1988 Apr;106(4):1171–1183. doi: 10.1083/jcb.106.4.1171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hirano T., Kinoshita N., Morikawa K., Yanagida M. Snap helix with knob and hole: essential repeats in S. pombe nuclear protein nuc2+. Cell. 1990 Jan 26;60(2):319–328. doi: 10.1016/0092-8674(90)90746-2. [DOI] [PubMed] [Google Scholar]
  11. Icho T., Wickner R. B. The MAK11 protein is essential for cell growth and replication of M double-stranded RNA and is apparently a membrane-associated protein. J Biol Chem. 1988 Jan 25;263(3):1467–1475. [PubMed] [Google Scholar]
  12. Keleher C. A., Redd M. J., Schultz J., Carlson M., Johnson A. D. Ssn6-Tup1 is a general repressor of transcription in yeast. Cell. 1992 Feb 21;68(4):709–719. doi: 10.1016/0092-8674(92)90146-4. [DOI] [PubMed] [Google Scholar]
  13. Koerner T. J., Hill J. E., Myers A. M., Tzagoloff A. High-expression vectors with multiple cloning sites for construction of trpE fusion genes: pATH vectors. Methods Enzymol. 1991;194:477–490. doi: 10.1016/0076-6879(91)94036-c. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Lockington R. A., Sealy-Lewis H. M., Scazzocchio C., Davies R. W. Cloning and characterization of the ethanol utilization regulon in Aspergillus nidulans. Gene. 1985;33(2):137–149. doi: 10.1016/0378-1119(85)90088-5. [DOI] [PubMed] [Google Scholar]
  16. Masuda H., Hirano T., Yanagida M., Cande W. Z. In vitro reactivation of spindle elongation in fission yeast nuc2 mutant cells. J Cell Biol. 1990 Feb;110(2):417–425. doi: 10.1083/jcb.110.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McIntosh J. R., Hering G. E. Spindle fiber action and chromosome movement. Annu Rev Cell Biol. 1991;7:403–426. doi: 10.1146/annurev.cb.07.110191.002155. [DOI] [PubMed] [Google Scholar]
  18. Meluh P. B., Rose M. D. KAR3, a kinesin-related gene required for yeast nuclear fusion. Cell. 1990 Mar 23;60(6):1029–1041. doi: 10.1016/0092-8674(90)90351-e. [DOI] [PubMed] [Google Scholar]
  19. Mitchison T. J. Polewards microtubule flux in the mitotic spindle: evidence from photoactivation of fluorescence. J Cell Biol. 1989 Aug;109(2):637–652. doi: 10.1083/jcb.109.2.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mitchison T. J., Salmon E. D. Poleward kinetochore fiber movement occurs during both metaphase and anaphase-A in newt lung cell mitosis. J Cell Biol. 1992 Nov;119(3):569–582. doi: 10.1083/jcb.119.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mitchison T. J., Sawin K. E. Tubulin flux in the mitotic spindle: where does it come from, where is it going? Cell Motil Cytoskeleton. 1990;16(2):93–98. doi: 10.1002/cm.970160202. [DOI] [PubMed] [Google Scholar]
  22. Mitchison T., Evans L., Schulze E., Kirschner M. Sites of microtubule assembly and disassembly in the mitotic spindle. Cell. 1986 May 23;45(4):515–527. doi: 10.1016/0092-8674(86)90283-7. [DOI] [PubMed] [Google Scholar]
  23. Morris N. R., Enos A. P. Mitotic gold in a mold: Aspergillus genetics and the biology of mitosis. Trends Genet. 1992 Jan;8(1):32–37. doi: 10.1016/0168-9525(92)90022-v. [DOI] [PubMed] [Google Scholar]
  24. Morris N. R. Mitotic mutants of Aspergillus nidulans. Genet Res. 1975 Dec;26(3):237–254. doi: 10.1017/s0016672300016049. [DOI] [PubMed] [Google Scholar]
  25. Murray A. W., Solomon M. J., Kirschner M. W. The role of cyclin synthesis and degradation in the control of maturation promoting factor activity. Nature. 1989 May 25;339(6222):280–286. doi: 10.1038/339280a0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. O'Donnell K. L., Osmani A. H., Osmani S. A., Morris N. R. bimA encodes a member of the tetratricopeptide repeat family of proteins and is required for the completion of mitosis in Aspergillus nidulans. J Cell Sci. 1991 Aug;99(Pt 4):711–719. doi: 10.1242/jcs.99.4.711. [DOI] [PubMed] [Google Scholar]
  28. Oakley B. R., Morris N. R. A beta-tubulin mutation in Aspergillus nidulans that blocks microtubule function without blocking assembly. Cell. 1981 Jun;24(3):837–845. doi: 10.1016/0092-8674(81)90109-4. [DOI] [PubMed] [Google Scholar]
  29. Oakley B. R., Oakley C. E., Yoon Y., Jung M. K. Gamma-tubulin is a component of the spindle pole body that is essential for microtubule function in Aspergillus nidulans. Cell. 1990 Jun 29;61(7):1289–1301. doi: 10.1016/0092-8674(90)90693-9. [DOI] [PubMed] [Google Scholar]
  30. Oakley C. E., Oakley B. R. Identification of gamma-tubulin, a new member of the tubulin superfamily encoded by mipA gene of Aspergillus nidulans. Nature. 1989 Apr 20;338(6217):662–664. doi: 10.1038/338662a0. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. PONTECORVO G., ROPER J. A., HEMMONS L. M., MACDONALD K. D., BUFTON A. W. J. The genetics of Aspergillus nidulans. Adv Genet. 1953;5:141–238. doi: 10.1016/s0065-2660(08)60408-3. [DOI] [PubMed] [Google Scholar]
  34. Rose M. D., Fink G. R. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast. Cell. 1987 Mar 27;48(6):1047–1060. doi: 10.1016/0092-8674(87)90712-4. [DOI] [PubMed] [Google Scholar]
  35. Rout M. P., Kilmartin J. V. Components of the yeast spindle and spindle pole body. J Cell Biol. 1990 Nov;111(5 Pt 1):1913–1927. doi: 10.1083/jcb.111.5.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rüther U., Müller-Hill B. Easy identification of cDNA clones. EMBO J. 1983;2(10):1791–1794. doi: 10.1002/j.1460-2075.1983.tb01659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sethi N., Monteagudo M. C., Koshland D., Hogan E., Burke D. J. The CDC20 gene product of Saccharomyces cerevisiae, a beta-transducin homolog, is required for a subset of microtubule-dependent cellular processes. Mol Cell Biol. 1991 Nov;11(11):5592–5602. doi: 10.1128/mcb.11.11.5592. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sikorski R. S., Boguski M. S., Goebl M., Hieter P. A repeating amino acid motif in CDC23 defines a family of proteins and a new relationship among genes required for mitosis and RNA synthesis. Cell. 1990 Jan 26;60(2):307–317. doi: 10.1016/0092-8674(90)90745-z. [DOI] [PubMed] [Google Scholar]
  39. Thomas J. H., Botstein D. A gene required for the separation of chromosomes on the spindle apparatus in yeast. Cell. 1986 Jan 17;44(1):65–76. doi: 10.1016/0092-8674(86)90485-x. [DOI] [PubMed] [Google Scholar]
  40. Vallen E. A., Hiller M. A., Scherson T. Y., Rose M. D. Separate domains of KAR1 mediate distinct functions in mitosis and nuclear fusion. J Cell Biol. 1992 Jun;117(6):1277–1287. doi: 10.1083/jcb.117.6.1277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Vallen E. A., Scherson T. Y., Roberts T., van Zee K., Rose M. D. Asymmetric mitotic segregation of the yeast spindle pole body. Cell. 1992 May 1;69(3):505–515. doi: 10.1016/0092-8674(92)90451-h. [DOI] [PubMed] [Google Scholar]
  42. 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]
  43. Weil C. F., Oakley C. E., Oakley B. R. Isolation of mip (microtubule-interacting protein) mutations of Aspergillus nidulans. Mol Cell Biol. 1986 Aug;6(8):2963–2968. doi: 10.1128/mcb.6.8.2963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Williams F. E., Varanasi U., Trumbly R. J. The CYC8 and TUP1 proteins involved in glucose repression in Saccharomyces cerevisiae are associated in a protein complex. Mol Cell Biol. 1991 Jun;11(6):3307–3316. doi: 10.1128/mcb.11.6.3307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Winey M., Goetsch L., Baum P., Byers B. MPS1 and MPS2: novel yeast genes defining distinct steps of spindle pole body duplication. J Cell Biol. 1991 Aug;114(4):745–754. doi: 10.1083/jcb.114.4.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Yelton M. M., Hamer J. E., Timberlake W. E. Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1470–1474. doi: 10.1073/pnas.81.5.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]

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