Abstract
Fodrin (nonerythroid spectrin) and its associated proteins have been previously implicated in the establishment of specialized membrane- cytoskeletal domains in differentiating cells. Using antiserum which is monospecific for the alpha-subunit of fodrin, we demonstrate that alpha- fodrin is present in oocytes and adult tissues of Xenopus laevis. Analyses of the de novo synthesis of alpha-fodrin during embryonic development reveal that alpha-fodrin is synthesized in oocytes, but not during early development. To investigate the level of control of alpha- fodrin expression, we isolated two cDNA clones for oocyte alpha-fodrin. The oocyte cDNA clones were identified as encoding portions of alpha- fodrin based on DNA sequence analysis and on the comparison of the predicted amino acid sequence of the cDNAs with the known sequence of human erythrocyte alpha-spectrin. The Xenopus alpha-fodrin cDNAs hybridize to a transcript of approximately 9 kb on RNA blots, and probably to a single gene type on genomic DNA blots. Both RNA blot analyses and S1 nuclease protection assays with the Xenopus alpha- fodrin cDNAs demonstrate that the observed decline in the de novo synthesis of alpha-fodrin polypeptides is controlled by a dramatic decrease in the abundance of alpha-fodrin transcripts after fertilization. In contrast, levels of actin transcripts do not decrease during this period. Inasmuch as steady-state levels of alpha-fodrin transcripts rise by the neurula stage of development, these results suggest that the synthesis of alpha-fodrin polypeptides during embryonic development of Xenopus is regulated, rather than constitutive, and that the primary level of control is the steady-state abundance of mRNA.
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- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bluemink J. G., Tertoolen L. G. The plasma-membrance IMP pattern as related to animal/vegetal polarity in the amphibian egg. Dev Biol. 1978 Feb;62(2):334–343. doi: 10.1016/0012-1606(78)90220-8. [DOI] [PubMed] [Google Scholar]
- Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
- Chou P. Y., Fasman G. D. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. doi: 10.1002/9780470122921.ch2. [DOI] [PubMed] [Google Scholar]
- Colot H. V., Rosbash M. Behavior of individual maternal pA+ RNAs during embryogenesis of Xenopus laevis. Dev Biol. 1982 Nov;94(1):79–86. doi: 10.1016/0012-1606(82)90070-7. [DOI] [PubMed] [Google Scholar]
- Dworkin M. B., Dawid I. B. Use of a cloned library for the study of abundant poly(A)+RNA during Xenopus laevis development. Dev Biol. 1980 May;76(2):449–464. doi: 10.1016/0012-1606(80)90393-0. [DOI] [PubMed] [Google Scholar]
- Dworkin M. B., Kay B. K., Hershey J. W., Dawid I. B. Mitochondrial RNAs are abundant in the poly(A)+RNA population of early frog embryos. Dev Biol. 1981 Sep;86(2):502–504. doi: 10.1016/0012-1606(81)90208-6. [DOI] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Franke W. W., Rathke P. C., Seib E., Trendelenburg M. F., Osborn M., Weber K. Distribution and mode of arrangement of microfilamentous structures and actin in the cortex of the amphibian oocyte. Cytobiologie. 1976 Dec;14(1):111–130. [PubMed] [Google Scholar]
- Franz J. K., Gall L., Williams M. A., Picheral B., Franke W. W. Intermediate-size filaments in a germ cell: Expression of cytokeratins in oocytes and eggs of the frog Xenopus. Proc Natl Acad Sci U S A. 1983 Oct;80(20):6254–6258. doi: 10.1073/pnas.80.20.6254. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Glenney J. R., Jr, Glenney P. Comparison of spectrin isolated from erythroid and non-erythroid sources. Eur J Biochem. 1984 Nov 2;144(3):529–539. doi: 10.1111/j.1432-1033.1984.tb08498.x. [DOI] [PubMed] [Google Scholar]
- Godsave S. F., Anderton B. H., Heasman J., Wylie C. C. Oocytes and early embryos of Xenopus laevis contain intermediate filaments which react with anti-mammalian vimentin antibodies. J Embryol Exp Morphol. 1984 Oct;83:169–187. [PubMed] [Google Scholar]
- Godsave S. F., Wylie C. C., Lane E. B., Anderton B. H. Intermediate filaments in the Xenopus oocyte: the appearance and distribution of cytokeratin-containing filaments. J Embryol Exp Morphol. 1984 Oct;83:157–167. [PubMed] [Google Scholar]
- Gurdon J. B., Wickens M. P. The use of Xenopus oocytes for the expression of cloned genes. Methods Enzymol. 1983;101:370–386. doi: 10.1016/0076-6879(83)01028-9. [DOI] [PubMed] [Google Scholar]
- Heikkila J. J., Kloc M., Bury J., Schultz G. A., Browder L. W. Acquisition of the heat-shock response and thermotolerance during early development of Xenopus laevis. Dev Biol. 1985 Feb;107(2):483–489. doi: 10.1016/0012-1606(85)90329-x. [DOI] [PubMed] [Google Scholar]
- Koenig E., Repasky E. A regional analysis of alpha-spectrin in the isolated Mauthner neuron and in isolated axons of the goldfish and rabbit. J Neurosci. 1985 Mar;5(3):705–714. doi: 10.1523/JNEUROSCI.05-03-00705.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Krieg P. A., Melton D. A. Developmental regulation of a gastrula-specific gene injected into fertilized Xenopus eggs. EMBO J. 1985 Dec 16;4(13A):3463–3471. doi: 10.1002/j.1460-2075.1985.tb04105.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lazarides E., Nelson W. J., Kasamatsu T. Segregation of two spectrin forms in the chicken optic system: a mechanism for establishing restricted membrane-cytoskeletal domains in neurons. Cell. 1984 Feb;36(2):269–278. doi: 10.1016/0092-8674(84)90220-4. [DOI] [PubMed] [Google Scholar]
- Levine J., Willard M. Fodrin: axonally transported polypeptides associated with the internal periphery of many cells. J Cell Biol. 1981 Sep;90(3):631–642. doi: 10.1083/jcb.90.3.631. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Levine J., Willard M. Redistribution of fodrin (a component of the cortical cytoplasm) accompanying capping of cell surface molecules. Proc Natl Acad Sci U S A. 1983 Jan;80(1):191–195. doi: 10.1073/pnas.80.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Minty A. J., Caravatti M., Robert B., Cohen A., Daubas P., Weydert A., Gros F., Buckingham M. E. Mouse actin messenger RNAs. Construction and characterization of a recombinant plasmid molecule containing a complementary DNA transcript of mouse alpha-actin mRNA. J Biol Chem. 1981 Jan 25;256(2):1008–1014. [PubMed] [Google Scholar]
- Moon R. T., Danilchik M. V., Hille M. B. An assessment of the masked message hypothesis: sea urchin egg messenger ribonucleoprotein complexes are efficient templates for in vitro protein synthesis. Dev Biol. 1982 Oct;93(2):389–403. doi: 10.1016/0012-1606(82)90126-9. [DOI] [PubMed] [Google Scholar]
- Moon R. T., Ngai J., Wold B. J., Lazarides E. Tissue-specific expression of distinct spectrin and ankyrin transcripts in erythroid and nonerythroid cells. J Cell Biol. 1985 Jan;100(1):152–160. doi: 10.1083/jcb.100.1.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Müller M. M., Carrasco A. E., DeRobertis E. M. A homeo-box-containing gene expressed during oogenesis in Xenopus. Cell. 1984 Nov;39(1):157–162. doi: 10.1016/0092-8674(84)90201-0. [DOI] [PubMed] [Google Scholar]
- Nelson W. J., Colaço C. A., Lazarides E. Involvement of spectrin in cell-surface receptor capping in lymphocytes. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1626–1630. doi: 10.1073/pnas.80.6.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Newport J., Kirschner M. A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982 Oct;30(3):675–686. doi: 10.1016/0092-8674(82)90272-0. [DOI] [PubMed] [Google Scholar]
- Rebagliati M. R., Weeks D. L., Harvey R. P., Melton D. A. Identification and cloning of localized maternal RNAs from Xenopus eggs. Cell. 1985 Oct;42(3):769–777. doi: 10.1016/0092-8674(85)90273-9. [DOI] [PubMed] [Google Scholar]
- Repasky E. A., Granger B. L., Lazarides E. Widespread occurrence of avian spectrin in nonerythroid cells. Cell. 1982 Jul;29(3):821–833. doi: 10.1016/0092-8674(82)90444-5. [DOI] [PubMed] [Google Scholar]
- Ruderman J. V., Woodland H. R., Sturgess E. A. Modulations of histone messenger RNA during the early development of Xenopus laevis. Dev Biol. 1979 Jul;71(1):71–82. doi: 10.1016/0012-1606(79)90083-6. [DOI] [PubMed] [Google Scholar]
- Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
- Schatten H., Cheney R., Balczon R., Willard M., Cline C., Simerly C., Schatten G. Localization of fodrin during fertilization and early development of sea urchins and mice. Dev Biol. 1986 Dec;118(2):457–466. doi: 10.1016/0012-1606(86)90016-3. [DOI] [PubMed] [Google Scholar]
- Sobel J. S., Alliegro M. A. Changes in the distribution of a spectrin-like protein during development of the preimplantation mouse embryo. J Cell Biol. 1985 Jan;100(1):333–336. doi: 10.1083/jcb.100.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Speicher D. W., Marchesi V. T. Erythrocyte spectrin is comprised of many homologous triple helical segments. Nature. 1984 Sep 13;311(5982):177–180. doi: 10.1038/311177a0. [DOI] [PubMed] [Google Scholar]
- Vacquier V. D. Dynamic changes of the egg cortex. Dev Biol. 1981 May;84(1):1–26. doi: 10.1016/0012-1606(81)90366-3. [DOI] [PubMed] [Google Scholar]
- Wasserman W. J., Richter J. D., Smith L. D. Protein synthesis during maturation promoting factor- and progesterone-induced maturation in Xenopus oocytes. Dev Biol. 1982 Jan;89(1):152–158. doi: 10.1016/0012-1606(82)90303-7. [DOI] [PubMed] [Google Scholar]