PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
 
Mol Cell Biol. 1991 September; 11(9): 4726–4731.
PMCID: PMC361369

A codon change in beta-tubulin which drastically affects microtubule structure in Drosophila melanogaster fails to produce a significant phenotype in Saccharomyces cerevisiae.

Abstract

The relative uniformity of microtubule ultrastructure in almost all eukaryotic cells is thought to be a consequence of the conserved elements of tubulin sequence. In support of this idea, a mutation in a beta-tubulin gene of Drosophila melanogaster, occurring at a highly conserved position, produces U-shaped microtubules, suggesting a defect in either nucleation or packing during assembly (M. T. Fuller, J. H. Caulton, J. A. Hutchens, T. C. Kaufman, and E. C. Raff, J. Cell Biol. 104:385-394, 1987, and J. E. Rudolph, M. Kimble, H. D. Hoyle, M. A. Subler, and E. C. Raff, Mol. Cell. Biol. 7:2231-2242, 1987). Surprisingly, we find that introducing the same mutation into the sole beta-tubulin gene of Saccharomyces cerevisiae has virtually no consequences for microtubule structure or function in that organism.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Boeke JD, LaCroute F, Fink GR. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. [PubMed]
  • Church GM, Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. [PubMed]
  • Cleveland DW, Sullivan KF. Molecular biology and genetics of tubulin. Annu Rev Biochem. 1985;54:331–365. [PubMed]
  • Das G, Hickey DR, Principio L, Conklin KT, Short J, Miller JR, McLendon G, Sherman F. Replacements of lysine 32 in yeast cytochrome c. Effects on the binding and reactivity with physiological partners. J Biol Chem. 1988 Dec 5;263(34):18290–18297. [PubMed]
  • Devlin PE, Ramachandran KL, Cate RL. Southern analysis of genomic DNA with unique and degenerate oligonucleotide probes: a method for reducing probe degeneracy. DNA. 1988 Sep;7(7):499–507. [PubMed]
  • Dinsmore JH, Sloboda RD. Microinjection of antibodies to a 62 kd mitotic apparatus protein arrests mitosis in dividing sea urchin embryos. Cell. 1989 Apr 7;57(1):127–134. [PubMed]
  • Fong D, Lee B. Beta tubulin gene of the parasitic protozoan Leishmania mexicana. Mol Biochem Parasitol. 1988 Oct;31(1):97–106. [PubMed]
  • Fuller MT, Caulton JH, Hutchens JA, Kaufman TC, Raff EC. Genetic analysis of microtubule structure: a beta-tubulin mutation causes the formation of aberrant microtubules in vivo and in vitro. J Cell Biol. 1987 Mar;104(3):385–394. [PMC free article] [PubMed]
  • Hall JL, Dudley L, Dobner PR, Lewis SA, Cowan NJ. Identification of two human beta-tubulin isotypes. Mol Cell Biol. 1983 May;3(5):854–862. [PMC free article] [PubMed]
  • Holm C, Meeks-Wagner DW, Fangman WL, Botstein D. A rapid, efficient method for isolating DNA from yeast. Gene. 1986;42(2):169–173. [PubMed]
  • Holzschu D, Principio L, Conklin KT, Hickey DR, Short J, Rao R, McLendon G, Sherman F. Replacement of the invariant lysine 77 by arginine in yeast iso-1-cytochrome c results in enhanced and normal activities in vitro and in vivo. J Biol Chem. 1987 May 25;262(15):7125–7131. [PubMed]
  • Huffaker TC, Thomas JH, Botstein D. Diverse effects of beta-tubulin mutations on microtubule formation and function. J Cell Biol. 1988 Jun;106(6):1997–2010. [PMC free article] [PubMed]
  • Katz WS, Solomon F. Diversity among beta-tubulins: a carboxy-terminal domain of yeast beta-tubulin is not essential in vivo. Mol Cell Biol. 1988 Jul;8(7):2730–2736. [PMC free article] [PubMed]
  • Katz W, Weinstein B, Solomon F. Regulation of tubulin levels and microtubule assembly in Saccharomyces cerevisiae: consequences of altered tubulin gene copy number. Mol Cell Biol. 1990 Oct;10(10):5286–5294. [PMC free article] [PubMed]
  • Kilmartin JV. Purification of yeast tubulin by self-assembly in vitro. Biochemistry. 1981 Jun 9;20(12):3629–3633. [PubMed]
  • Neff NF, Thomas JH, Grisafi P, Botstein D. Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo. Cell. 1983 May;33(1):211–219. [PubMed]
  • Pallas D, Solomon F. Cytoplasmic microtubule-associated proteins: phosphorylation at novel sites is correlated with their incorporation into assembled microtubules. Cell. 1982 Sep;30(2):407–414. [PubMed]
  • Reed KC, Mann DA. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. [PMC free article] [PubMed]
  • Rudolph JE, Kimble M, Hoyle HD, Subler MA, Raff EC. Three Drosophila beta-tubulin sequences: a developmentally regulated isoform (beta 3), the testis-specific isoform (beta 2), and an assembly-defective mutation of the testis-specific isoform (B2t8) reveal both an ancient divergence in metazoan isotypes and structural constraints for beta-tubulin function. Mol Cell Biol. 1987 Jun;7(6):2231–2242. [PMC free article] [PubMed]
  • Schatz PJ, Solomon F, Botstein D. Isolation and characterization of conditional-lethal mutations in the TUB1 alpha-tubulin gene of the yeast Saccharomyces cerevisiae. Genetics. 1988 Nov;120(3):681–695. [PubMed]
  • Sullivan KF, Machlin PS, Ratrie H, 3rd, Cleveland DW. Sequence and expression of the chicken beta 3 tubulin gene. A vertebrate testis beta-tubulin isotype. J Biol Chem. 1986 Oct 5;261(28):13317–13322. [PubMed]
  • Thomas JH, Neff NF, Botstein D. Isolation and characterization of mutations in the beta-tubulin gene of Saccharomyces cerevisiae. Genetics. 1985 Dec;111(4):715–734. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)