Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1983 Apr 1;96(4):1056–1063. doi: 10.1083/jcb.96.4.1056

Multiple forms of tubulin in polytomella and chlamydomonas: evidence for a precursor of flagellar α-tubulin

TW McKeithan, PA Lefebvre, CD Silflow, JL Rosenbaum
PMCID: PMC2112316  PMID: 6833391

Abstract

The quadriflagellate alga polytomella agilis contains several α-tubulins with distinct isoelectric points (McKeithan, T.W., and J.L. Rosenbaum, 1981, J. Cell Biol., 91:352-360). While α-3 is the major component in flagella, α-1 predominates in cytoskeletal microtubules. For determination of whether the differences in α- tubulins are due to distinct genes or to posttranslational modification of a common α-tubulin precursor, poly A+ RNA was isolated from deflagellated and control (nonregenerating) cells and translated in vitro. Approximately twice as much α-1 was synthesized using RNA from deflagellated as compared to control cells; however, there was no detectable synthesis in vitro of α-3 in either. These results suggest that α -3 tubulin is formed in vivo by posttranslational modification of a form co- migrating with, and possibly identical to, cytoskeletal α-tubulin. In the related alga chlamydomonas reinhardii deflagellation greatly stimulates synthesis of tubulin and tubulin mRNA. As in polytomella, the principal α-tubulin synthesized both in vivo and in vitro following deflagellation in chlamydomonas is more basic than the major α-tubulin and appears to correspond to α-1 tubulin in polytomella. The conversion of α-1 to α-3 receives additional support from in vivo labeling and pulse-chase experiments. In addition, in both polytomella and chlamydomonas some conversion of α-1 to α-3 appears to occur even when protein synthesis is inhibited.

Full Text

The Full Text of this article is available as a PDF (2.0 MB).

Selected References

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

  1. Arce C. A., Rodriguez J. A., Barra H. S., Caputo R. Incorporation of L-tyrosine, L-phenylalanine and L-3,4-dihydroxyphenylalanine as single units into rat brain tubulin. Eur J Biochem. 1975 Nov 1;59(1):145–149. doi: 10.1111/j.1432-1033.1975.tb02435.x. [DOI] [PubMed] [Google Scholar]
  2. Argaraña C. E., Arce C. A., Barra H. S., Caputto R. In vivo incorporation of [14C]tyrosine into the C-terminal position of the alpha subunit of tubulin. Arch Biochem Biophys. 1977 Apr 30;180(2):264–268. doi: 10.1016/0003-9861(77)90037-6. [DOI] [PubMed] [Google Scholar]
  3. Barra H. S., Arce C. A., Rodríguez J. A., Caputto R. Some common properties of the protein that incorporates tyrosine as a single unit and the microtubule proteins. Biochem Biophys Res Commun. 1974 Oct 23;60(4):1384–1390. doi: 10.1016/0006-291x(74)90351-9. [DOI] [PubMed] [Google Scholar]
  4. Bloodgood R. A. Resorption of organelles containing microtubules. Cytobios. 1974 Mar-Apr;9(35):142–161. [PubMed] [Google Scholar]
  5. Brown D. L., Rogers K. A. Hydrostatic pressure-induced internalization of flagellar axonemes, disassembly, and reutilization during flagellar regeneration in Polytomella. Exp Cell Res. 1978 Dec;117(2):313–324. doi: 10.1016/0014-4827(78)90145-3. [DOI] [PubMed] [Google Scholar]
  6. Brunke K. J., Young E. E., Buchbinder B. U., Weeks D. P. Coordinate regulation of the four tubulin genes of Chlamydomonas reinhardi. Nucleic Acids Res. 1982 Feb 25;10(4):1295–1310. doi: 10.1093/nar/10.4.1295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eipper B. A. Rat brain tubulin and protein kinase activity. J Biol Chem. 1974 Mar 10;249(5):1398–1406. [PubMed] [Google Scholar]
  8. Hallak M. E., Rodriguez J. A., Barra H. S., Caputto R. Release of tyrosine from tyrosinated tubulin. Some common factors that affect this process and the assembly of tubulin. FEBS Lett. 1977 Feb 1;73(2):147–150. doi: 10.1016/0014-5793(77)80968-x. [DOI] [PubMed] [Google Scholar]
  9. Kemphues K. J., Raff E. C., Raff R. A., Kaufman T. C. Mutation in a testis-specific beta-tubulin in Drosophila: analysis of its effects on meiosis and map location of the gene. Cell. 1980 Sep;21(2):445–451. doi: 10.1016/0092-8674(80)90481-x. [DOI] [PubMed] [Google Scholar]
  10. Kemphues K. J., Raff R. A., Kaufman T. C., Raff E. C. Mutation in a structural gene for a beta-tubulin specific to testis in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3991–3995. doi: 10.1073/pnas.76.8.3991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kowit J. D., Fulton C. Programmed synthesis of tubulin for the flagella that develop during cell differentiation in Naegleria gruberi. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2877–2881. doi: 10.1073/pnas.71.7.2877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kowit J. D., Fulton C. Purification and properties of flagellar outer doublet tubulin from Naegleria gruberi and a radioimmune assay for tubulin. J Biol Chem. 1974 Jun 10;249(11):3638–3646. [PubMed] [Google Scholar]
  13. Kumar N., Flavin M. Preferential action of a brain detyrosinolating carboxypeptidase on polymerized tubulin. J Biol Chem. 1981 Jul 25;256(14):7678–7686. [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lefebvre P. A., Nordstrom S. A., Moulder J. E., Rosenbaum J. L. Flagellar elongation and shortening in Chlamydomonas. IV. Effects of flagellar detachment, regeneration, and resorption on the induction of flagellar protein synthesis. J Cell Biol. 1978 Jul;78(1):8–27. doi: 10.1083/jcb.78.1.8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lefebvre P. A., Silflow C. D., Wieben E. D., Rosenbaum J. L. Increased levels of mRNAs for tubulin and other flagellar proteins after amputation or shortening of Chlamydomonas flagella. Cell. 1980 Jun;20(2):469–477. doi: 10.1016/0092-8674(80)90633-9. [DOI] [PubMed] [Google Scholar]
  17. McKeithan T. W., Rosenbaum J. L. Multiple forms of tubulin in the cytoskeletal and flagellar microtubules of Polytomella. J Cell Biol. 1981 Nov;91(2 Pt 1):352–360. doi: 10.1083/jcb.91.2.352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Raybin D., Flavin M. An enzyme tyrosylating alpha-tubulin and its role in microtubule assembly. Biochem Biophys Res Commun. 1975 Aug 4;65(3):1088–1095. doi: 10.1016/s0006-291x(75)80497-9. [DOI] [PubMed] [Google Scholar]
  20. Rosenbaum J. L., Child F. M. Flagellar regeneration in protozoan flagellates. J Cell Biol. 1967 Jul;34(1):345–364. doi: 10.1083/jcb.34.1.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rosenbaum J. L., Moulder J. E., Ringo D. L. Flagellar elongation and shortening in Chlamydomonas. The use of cycloheximide and colchicine to study the synthesis and assembly of flagellar proteins. J Cell Biol. 1969 May;41(2):600–619. doi: 10.1083/jcb.41.2.600. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sheir-Neiss G., Lai M. H., Morris N. R. Identification of a gene for beta-tubulin in Aspergillus nidulans. Cell. 1978 Oct;15(2):639–647. doi: 10.1016/0092-8674(78)90032-6. [DOI] [PubMed] [Google Scholar]
  23. Silflow C. D., Rosenbaum J. L. Multiple alpha- and beta-tubulin genes in Chlamydomonas and regulation of tubulin mRNA levels after deflagellation. Cell. 1981 Apr;24(1):81–88. doi: 10.1016/0092-8674(81)90503-1. [DOI] [PubMed] [Google Scholar]
  24. Stephens R. E. Primary structural differences among tubulin subunits from flagella, cilia, and the cytoplasm. Biochemistry. 1978 Jul 11;17(14):2882–2891. doi: 10.1021/bi00607a029. [DOI] [PubMed] [Google Scholar]
  25. Stephens R. E. Structural chemistry of the axoneme: evidence for chemically and functionally unique tubulin dimers in outer fibers. Soc Gen Physiol Ser. 1975;30:181–206. [PubMed] [Google Scholar]
  26. Stephens R. E. Thermal fractionation of outer fiber doublet microtubules into A- and B-subfiber components. A- and B-tubulin. J Mol Biol. 1970 Feb 14;47(3):353–363. doi: 10.1016/0022-2836(70)90307-4. [DOI] [PubMed] [Google Scholar]
  27. Weeks D. P., Collis P. S. Induction of microtubule protein synthesis in Chlamydomonas reinhardi during flagellar regeneration. Cell. 1976 Sep;9(1):15–27. doi: 10.1016/0092-8674(76)90048-9. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES