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. 1988 Jun;8(6):2280–2287. doi: 10.1128/mcb.8.6.2280

Transcriptional regulation of coordinate changes in flagellar mRNAs during differentiation of Naegleria gruberi amebae into flagellates.

J H Lee 1, C J Walsh 1
PMCID: PMC363424  PMID: 3405205

Abstract

The nuclear run-on technique was used to measure the rate of transcription of flagellar genes during the differentiation of Naegleria gruberi amebae into flagellates. Synthesis of mRNAs for the axonemal proteins alpha- and beta-tubulin and flagellar calmodulin, as well as a coordinately regulated poly(A)+ RNA that codes for an unidentified protein, showed transient increases averaging 22-fold. The rate of synthesis of two poly(A)+ RNAs common to amebae and flagellates was low until the transcription of the flagellar genes began to decline, at which time synthesis of the RNAs found in amebae increased 3- to 10-fold. The observed changes in the rate of transcription can account quantitatively for the 20-fold increase in flagellar mRNA concentration during the differentiation. The data for the flagellar calmodulin gene demonstrate transcriptional regulation for a nontubulin axonemal protein. The data also demonstrate at least two programs of transcriptional regulation during the differentiation and raise the intriguing possibility that some significant fraction of the nearly 200 different proteins of the flagellar axoneme is transcriptionally regulated during the 1 h it takes N. gruberi amebae to form visible flagella.

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Selected References

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

  1. Baker E. J., Keller L. R., Schloss J. A., Rosenbaum J. L. Protein synthesis is required for rapid degradation of tubulin mRNA and other deflagellation-induced RNAs in Chlamydomonas reinhardi. Mol Cell Biol. 1986 Jan;6(1):54–61. doi: 10.1128/mcb.6.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker E. J., Schloss J. A., Rosenbaum J. L. Rapid changes in tubulin RNA synthesis and stability induced by deflagellation in Chlamydomonas. J Cell Biol. 1984 Dec;99(6):2074–2081. doi: 10.1083/jcb.99.6.2074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brunke K. J., Anthony J. G., Sternberg E. J., Weeks D. P. Repeated consensus sequence and pseudopromoters in the four coordinately regulated tubulin genes of Chlamydomonas reinhardi. Mol Cell Biol. 1984 Jun;4(6):1115–1124. doi: 10.1128/mcb.4.6.1115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Caron J. M., Jones A. L., Kirschner M. W. Autoregulation of tubulin synthesis in hepatocytes and fibroblasts. J Cell Biol. 1985 Nov;101(5 Pt 1):1763–1772. doi: 10.1083/jcb.101.5.1763. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cleveland D. W., Havercroft J. C. Is apparent autoregulatory control of tubulin synthesis nontranscriptionally regulated? J Cell Biol. 1983 Sep;97(3):919–924. doi: 10.1083/jcb.97.3.919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cleveland D. W., Lopata M. A., Sherline P., Kirschner M. W. Unpolymerized tubulin modulates the level of tubulin mRNAs. Cell. 1981 Aug;25(2):537–546. doi: 10.1016/0092-8674(81)90072-6. [DOI] [PubMed] [Google Scholar]
  8. Dingle A. D., Fulton C. Development of the flagellar apparatus of Naegleria. J Cell Biol. 1966 Oct;31(1):43–54. doi: 10.1083/jcb.31.1.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fulton C. Cell differentiation in Naegleria gruberi. Annu Rev Microbiol. 1977;31:597–629. doi: 10.1146/annurev.mi.31.100177.003121. [DOI] [PubMed] [Google Scholar]
  10. Fulton C., Cheng K. L., Lai E. Y. Two calmodulins in Naegleria flagellates: characterization, intracellular segregation, and programmed regulation of mRNA abundance during differentiation. J Cell Biol. 1986 May;102(5):1671–1678. doi: 10.1083/jcb.102.5.1671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fulton C., Dingle A. D. Appearance of the flagellate phenotype in populations of Naegleria amebae. Dev Biol. 1967 Feb;15(2):165–191. doi: 10.1016/0012-1606(67)90012-7. [DOI] [PubMed] [Google Scholar]
  12. Fulton C., Dingle A. D. Basal bodies, but not centrioles, in Naegleria. J Cell Biol. 1971 Dec;51(3):826–836. doi: 10.1083/jcb.51.3.826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fulton C., Walsh C. Cell differentiation and flagellar elongation in Naegleria gruberi. Dependence on transcription and translation. J Cell Biol. 1980 May;85(2):346–360. doi: 10.1083/jcb.85.2.346. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gilroy T. E., Beaudet A. L., Yu J. A method for analyzing transcription using permeabilized cells. Anal Biochem. 1984 Dec;143(2):350–360. doi: 10.1016/0003-2697(84)90674-2. [DOI] [PubMed] [Google Scholar]
  15. Keller L. R., Schloss J. A., Silflow C. D., Rosenbaum J. L. Transcription of alpha- and beta-tubulin genes in vitro in isolated Chlamydomonas reinhardi nuclei. J Cell Biol. 1984 Mar;98(3):1138–1143. doi: 10.1083/jcb.98.3.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Lai E. Y., Walsh C., Wardell D., Fulton C. Programmed appearance of translatable flagellar tubulin mRNA during cell differentiation in Naegleria. Cell. 1979 Aug;17(4):867–878. doi: 10.1016/0092-8674(79)90327-1. [DOI] [PubMed] [Google Scholar]
  19. Larson D. E., Dingle A. D. Development of the flagellar rootlet during Naegleria flagellate differentiation. Dev Biol. 1981 Aug;86(1):227–235. doi: 10.1016/0012-1606(81)90334-1. [DOI] [PubMed] [Google Scholar]
  20. Larson D. E., Dingle A. D. Isolation, ultrastructure, and protein composition of the flagellar rootlet of Naegleria gruberi. J Cell Biol. 1981 Jun;89(3):424–432. doi: 10.1083/jcb.89.3.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lau J. T., Pittenger M. F., Cleveland D. W. Reconstruction of appropriate tubulin and actin gene regulation after transient transfection of cloned beta-tubulin and beta-actin genes. Mol Cell Biol. 1985 Jul;5(7):1611–1620. doi: 10.1128/mcb.5.7.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Luck D. J., Huang B., Piperno G. Genetic and biochemical analysis of the eukaryotic flagellum. Symp Soc Exp Biol. 1982;35:399–419. [PubMed] [Google Scholar]
  24. Mar J., Lee J. H., Shea D., Walsh C. J. New poly(A)+RNAs appear coordinately during the differentiation of Naegleria gruberi amebae into flagellates. J Cell Biol. 1986 Feb;102(2):353–361. doi: 10.1083/jcb.102.2.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. McKnight G. S., Palmiter R. D. Transcriptional regulation of the ovalbumin and conalbumin genes by steroid hormones in chick oviduct. J Biol Chem. 1979 Sep 25;254(18):9050–9058. [PubMed] [Google Scholar]
  26. Remillard S. P., Witman G. B. Synthesis, transport, and utilization of specific flagellar proteins during flagellar regeneration in Chlamydomonas. J Cell Biol. 1982 Jun;93(3):615–631. doi: 10.1083/jcb.93.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Shea D. K., Walsh C. J. mRNAs for alpha- and beta-tubulin and flagellar calmodulin are among those coordinately regulated when Naegleria gruberi amebae differentiate into flagellates. J Cell Biol. 1987 Sep;105(3):1303–1309. doi: 10.1083/jcb.105.3.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Walsh C., Fulton C. Transcription during cell differentiation in Naegleria gruberi. Preferential synthesis of messenger RNA. Biochim Biophys Acta. 1973 Jun 8;312(1):52–71. doi: 10.1016/0005-2787(73)90052-x. [DOI] [PubMed] [Google Scholar]
  30. Walsh C. Synthesis and assembly of the cytoskeleton of Naegleria gruberi flagellates. J Cell Biol. 1984 Feb;98(2):449–456. doi: 10.1083/jcb.98.2.449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Youngblom J., Schloss J. A., Silflow C. D. The two beta-tubulin genes of Chlamydomonas reinhardtii code for identical proteins. Mol Cell Biol. 1984 Dec;4(12):2686–2696. doi: 10.1128/mcb.4.12.2686. [DOI] [PMC free article] [PubMed] [Google Scholar]

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