Abstract
Deciliation by hypertonic shock of embryos of the sea urchin Lytechinus pictus resulted in an increase in synthesis of alpha- and beta-tubulins, the consequence of an increased concentration of RNA encoding the tubulins. RNA run-on assays in isolated nuclei indicated that this response is due to a transient increase in the rate of synthesis of tubulin RNA beginning within 5 min of deciliation. This enhancement of tubulin gene transcription also occurred in deciliated embryos treated with the microtubule-depolymerizing agent colcemid; thus the reaction to deciliation is not a response to a reduction in concentration of unpolymerized tubulin utilized for ciliogenesis. In deciliated embryos treated with colcemid, the elevated level of tubulin RNA declined rapidly, due to its destabilization by the elevated concentration of unpolymerized tubulin. The increased transcription of tubulin genes is a response to the loss of cilia, not to the hypertonic shock, and occurs even when cilium regeneration is prevented. Inhibition of protein synthesis with puromycin or emetine did not prevent the transcriptional enhancement but stabilized tubulin mRNA, resulting in increased accumulation of tubulin mRNA after deciliation.
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- Alexandraki D., Ruderman J. V. Evolution of alpha q- and beta-tubulin genes as inferred by the nucleotide sequences of sea urchin cDNA clones. J Mol Evol. 1983;19(6):397–410. doi: 10.1007/BF02102315. [DOI] [PubMed] [Google Scholar]
- Alexandraki D., Ruderman J. V. Expression of alpha- and beta-tubulin genes during development of sea urchin embryos. Dev Biol. 1985 Jun;109(2):436–451. doi: 10.1016/0012-1606(85)90470-1. [DOI] [PubMed] [Google Scholar]
- Alexandraki D., Ruderman J. V. Multiple polymorphic alpha- and beta-tubulin mRNAs are present in sea urchin eggs. Proc Natl Acad Sci U S A. 1985 Jan;82(1):134–138. doi: 10.1073/pnas.82.1.134. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Aronson J., Inoué S. Reversal by light of the action of N-methyl N-desacetyl colchicine on mitosis. J Cell Biol. 1970 May;45(2):470–477. doi: 10.1083/jcb.45.2.470. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Auclair W., Siegel B. W. Cilia regeneration in the sea urchin embryo: evidence for a pool of ciliary proteins. Science. 1966 Nov 18;154(3751):913–915. doi: 10.1126/science.154.3751.913. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Ben-Ze'ev A., Farmer S. R., Penman S. Mechanisms of regulating tubulin synthesis in cultured mammalian cells. Cell. 1979 Jun;17(2):319–325. doi: 10.1016/0092-8674(79)90157-0. [DOI] [PubMed] [Google Scholar]
- Bibring T., Baxandall J. Tubulin synthesis in sea urchin embryos. II. Ciliary A tubulin derives from the unfertilized egg. Dev Biol. 1981 Apr 15;83(1):122–126. doi: 10.1016/s0012-1606(81)80014-0. [DOI] [PubMed] [Google Scholar]
- Bibring T., Baxandall J. Tubulin synthesis in sea urchin embryos: almost all tubulin of the first cleavage mitotic apparatus derives from the unfertilized egg. Dev Biol. 1977 Jan;55(1):191–195. doi: 10.1016/0012-1606(77)90330-x. [DOI] [PubMed] [Google Scholar]
- Burns R. G. Kinetics of the regeneration of sea-urchin cilia. J Cell Sci. 1973 Jul;13(1):55–67. doi: 10.1242/jcs.13.1.55. [DOI] [PubMed] [Google Scholar]
- Bédard P. A., Brandhorst B. P. Patterns of protein synthesis and metabolism during sea urchin embryogenesis. Dev Biol. 1983 Mar;96(1):74–83. doi: 10.1016/0012-1606(83)90312-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Caron J. M., Jones A. L., Rall L. B., Kirschner M. W. Autoregulation of tubulin synthesis in enucleated cells. Nature. 1985 Oct 17;317(6038):648–651. doi: 10.1038/317648a0. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Cleveland D. W., Pittenger M. F., Feramisco J. R. Elevation of tubulin levels by microinjection suppresses new tubulin synthesis. Nature. 1983 Oct 20;305(5936):738–740. doi: 10.1038/305738a0. [DOI] [PubMed] [Google Scholar]
- Conlon R. A., Tufaro F., Brandhorst B. P. Post-transcriptional restriction of gene expression in sea urchin interspecies hybrid embryos. Genes Dev. 1987 Jun;1(4):337–346. doi: 10.1101/gad.1.4.337. [DOI] [PubMed] [Google Scholar]
- GROSS P. R., COUSINEAU G. H. MACROMOLECULE SYNTHESIS AND THE INFLUENCE OF ACTINOMYCIN ON EARLY DEVELOPMENT. Exp Cell Res. 1964 Feb;33:368–395. doi: 10.1016/0014-4827(64)90002-3. [DOI] [PubMed] [Google Scholar]
- Greenhouse G. A., Hynes R. O., Gross P. R. Sea urchin embryos are permeable to actinomycin. Science. 1971 Feb 19;171(3972):686–689. doi: 10.1126/science.171.3972.686. [DOI] [PubMed] [Google Scholar]
- Groudine M., Peretz M., Weintraub H. Transcriptional regulation of hemoglobin switching in chicken embryos. Mol Cell Biol. 1981 Mar;1(3):281–288. doi: 10.1128/mcb.1.3.281. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Harlow P., Nemer M. Developmental and tissue-specific regulation of beta-tubulin gene expression in the embryo of the sea urchin Strongylocentrotus purpuratus. Genes Dev. 1987 Apr;1(2):147–160. doi: 10.1101/gad.1.2.147. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Lefebvre P. A., Rosenbaum J. L. Regulation of the synthesis and assembly of ciliary and flagellar proteins during regeneration. Annu Rev Cell Biol. 1986;2:517–546. doi: 10.1146/annurev.cb.02.110186.002505. [DOI] [PubMed] [Google Scholar]
- Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
- Merlino G. T., Chamberlain J. P., Kleinsmith L. J. Effects of deciliation of tubulin messenger RNA activity in sea urchin embryos. J Biol Chem. 1978 Oct 10;253(19):7078–7085. [PubMed] [Google Scholar]
- Morris G. F., Marzluff W. F. A factor in sea urchin eggs inhibits transcription in isolated nuclei by sea urchin RNA polymerase III. Biochemistry. 1983 Feb 1;22(3):645–653. doi: 10.1021/bi00272a019. [DOI] [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- Pittenger M. F., Cleveland D. W. Retention of autoregulatory control of tubulin synthesis in cytoplasts: demonstration of a cytoplasmic mechanism that regulates the level of tubulin expression. J Cell Biol. 1985 Nov;101(5 Pt 1):1941–1952. doi: 10.1083/jcb.101.5.1941. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Raff R. A., Greenhouse G., Gross K. W., Gross P. R. Synthesis and storage of microtubule proteins by sea urchin embryos. J Cell Biol. 1971 Aug;50(2):516–527. doi: 10.1083/jcb.50.2.516. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Schloss J. A., Silflow C. D., Rosenbaum J. L. mRNA abundance changes during flagellar regeneration in Chlamydomonas reinhardtii. Mol Cell Biol. 1984 Mar;4(3):424–434. doi: 10.1128/mcb.4.3.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stephens R. E. Differential protein synthesis and utilization during cilia formation in sea urchin embryos. Dev Biol. 1977 Dec;61(2):311–329. doi: 10.1016/0012-1606(77)90301-3. [DOI] [PubMed] [Google Scholar]
- Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Weeks D. P., Collis P., Gealt M. A. Control of induction of tubulin synthesis in Chlamydomonas reinhardi. Nature. 1977 Aug 18;268(5621):667–668. doi: 10.1038/268667a0. [DOI] [PubMed] [Google Scholar]