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Plant Molecular Biology

, Volume 46, Issue 1, pp 79–88 | Cite as

Identification of a cytoskeleton-associated 120 kDa RNA-binding protein in developing rice seeds

  • R. Sami-Subbu
  • Sang-Bong Choi
  • Yujia Wu
  • Changlin Wang
  • Thomas W. Okita
Article

Abstract

During rice seed development, prolamine RNAs are localized to the surface of the prolamine storage protein bodies (PBs), organelles bounded by the endoplasmic reticulum (ER). The exact mechanism by which prolamine RNAs are enriched on this ER subdomain is not known but recent evidence indicates the directed transport and targeting of prolamine RNAs to the prolamine PBs. As such a process involves RNA signal determinants and cytoskeleton-interacting proteins that recognize these signals, we obtained an enriched cytoskeleton-PB fraction and identified a prominent RNA-binding activity, Rp120, by RNA-binding UV-cross-linking assay. Recombinant cDNA clones of Rp120 revealed that the primary sequence shared considerable structural homology to the human transcriptional coactivator p100 and possessed a modular organization, four nucleic acid-binding SN domains, a tudor domain and a coil-coil domain. Consistent with the presence of SN domains, Rp120 binds a variety of RNAs including prolamine RNA. Interaction with the latter RNA, however, was specific as binding activity was evident only to the prolamine 3′ UTR and not to the 5′ UTR or coding sequences. Rp120 is also able to interact with other proteins as its sedimentation behavior in sucrose density gradient suggests an association with the cytoskeleton. The presence of a tudor domain, suggested to have a role in RNA processing or transport, together with the SN and coiled-coil domains are consistent with the view that Rp120 may be involved in RNA sorting in rice endosperm.

cytoskeleton protein bodies RNA-binding protein rice prolamine Rp120 

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References

  1. Abe, S. and Davies, E. 1995. Methods for isolation and analysis of the cytoskeleton. Meth. Cell Biol. 50: 223–236.Google Scholar
  2. Ainger, K., Avossa, D., Diana, A.S., Barry, C., Barbarese, E. and Carson, J.H. 1997. Transport and localization elements in myelin basic protein mRNA. J. Cell Biol. 138: 1077–1087.Google Scholar
  3. Callebaut, I. and Mornon, J.P. 1997. The human EBNA-2 coactivator p100: multidomain organization and realtionship to the staphylococcus nuclease fold and to the tudor protein involved in Drosophila melanogaster development. Biochem. J. 321: 125–132.Google Scholar
  4. Choi, S.-B., Wang, C., Muench, D.G., Ozawa, K., Franceschi, V.R., Wu, Y. and Okita, T.W. 2000. Messenger RNA targeting of rice seed storage proteins to specific ER subdomains. Nature 407: 765–767.Google Scholar
  5. Davies, E., Fillingham, B.D., Oto, Y. and Abe, S. 1991. Evidence for the existence of cytoskeleton-bound polysomes in plants. Cell Biol. Int. Rep. 15: 973–981.Google Scholar
  6. Davies, E., Comer, E.C., Lionberger, J.M., Stankovic, B. and Abe, S. 1993. Cytoskeleton-bound polysomes in plants. III. Polysome-cytoskeleton-membrane interactions in maize. Cell Biol. Int. 17: 331–340.Google Scholar
  7. Davies, E., Fillingham, B.D. and Abe, S. 1996. The plant cytoskeleton. In: J.E. Hesketh and I.F. Pryme (Eds.) The Cytoskeleton, JAI Press, Greenwich, CT, pp. 405–409.Google Scholar
  8. Deshler, J.O., Highett, M.I., Abramson, T. and Schnapp, B.J. 1998. A highly conserved RNA-binding protein for cytoplasmic mRNA localization in vertebrates. Curr. Biol. 8: 489–496.Google Scholar
  9. Gillespie, D.E. and Berg, C.A. 1995. Homeless is required for RNA localization in Drosophila oogenesis and encodes a new member of the DE-H family of RNA-dependent ATPases. Genes Dev. 9: 2495–2508.Google Scholar
  10. Golumbeski, G.S., Bardsley, A., Tax, F. and Boswell, R.E. 1991. tudor, a posterior-group gene of Drosophila melanogaster, encodes a novel protein and an mRNA localized during mid-oogenesis. Genes Dev. 5: 2495–2508.Google Scholar
  11. Hesketh, J.E. 1996. Sorting of messenger RNAs in the cytoplasm: mRNA localization and the cytoskeleton. Exp. Cell Res. 225: 219–236.Google Scholar
  12. Kim, W.T. and Okita, T.W. 1988. Structure, expression, and heterogeneity of the rice seed prolamines. Plant Physiol. 88: 649–655.Google Scholar
  13. Kim, W.T., Li, X. and Okita, T.W. 1993. Expression of storage protein multigene families in developing rice endosperm. Plant Cell Physiol. 34: 595–603.Google Scholar
  14. Lefebvre, S., Bürglen, L., Reboullet, S., Clermont, O., Burlet, P., Viollet, L., Benichou, B., Cruaud, C., Millasseau, P., Zeviani, M., Le Paslier, D., Frézal, J., Cohen, D., Weissenbach, J., Munnich, A. and Melki, J. 1995. Identification and characterization of a spinal muscular atrophy-determining gene. Cell 13: 155–165.Google Scholar
  15. Li, X., Franceschi, V.R. and Okita, T.W. 1993. Segregation of storage protein mRNAs on the rough endoplasmic reticulum membranes of rice endosperm cells. Cell 72: 869–879.Google Scholar
  16. Macdonald, P.M. and Kerr, K. 1997. Redundant RNA recognition events in bicoid mRNA localization. RNA 3: 1413–1420.Google Scholar
  17. Muench, D.G. and Okita, T.W. 1997. The storage proteins of rice and oat. In: B.A. Larkins and I.K. Vasil (Eds.) Cellular and Molecular Biology of Plant Development, Kluwer Academic Publishers. Dordrecht, Netherlands, pp. 281–330.Google Scholar
  18. Muench, D.G., Wu, Y., Coughlan, S.J. and Okita, T.W. 1998. Evidence for a cytoskeleton-associated binding site in prolamine mRNA localization to the protein bodies in rice endosperm. Plant Physiol. 116: 559–569.Google Scholar
  19. Muench, D.G., Ogawa, M. and Okita, T.W. 1999. The prolamins of rice. In: P.R. Shewry and R. Casey (Eds.) Seed Proteins, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 93–108.Google Scholar
  20. Muench, D.G., Chuong, S.D.X., Franceschi, V.R. and Okita, T.W. 2000. Developing prolamine protein bodies are associated with the cortical cytoskeleton in rice endosperm cells. Planta, in press.Google Scholar
  21. Murzin, A.G. 1993. OB (oligonucleotide/oligosaccharide binding)-fold: common structural and functional solution for non-homologous sequences. EMBO J. 12: 861–867.Google Scholar
  22. Okita, T.W. and Rogers, J.C. 1996. Compartmentation of proteins in the endomembrane system of plant cells. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 327–350.Google Scholar
  23. Okita, T.W., Hwang, Y.S., Hnilo, J., Kim, W.T., Aryan, A.P., Larsen, R. and Krishnan, H.B. 1989. Structure and expression of the rice glutelin multigene family. J. Biol. Chem. 264: 12573–12581.Google Scholar
  24. Okita, T.W., Li, X. and Roberts, M.W. 1994. Targeting of mRNAs to domains of the endoplasmic reticulum. Trends Cell Biol. 4: 91–96.Google Scholar
  25. Ponting, C.P. 1997. Tudor domains in proteins that interact with RNA. Trends Biochem. Sci. 22: 51–52.Google Scholar
  26. Raker, B.D., Luhrmann, V. and Fischer, U. 1999. Essential role for the tudor domain of SMN in spliceosomal U snRNP assembly: implications for spinal muscular atrophy. Hum. Mol. Genet. 8: 2351–2357.Google Scholar
  27. Sami-Subbu, R., Muench, D.G. and Okita, T.W. 1999. A cytoskeleton-protein body associated RNA-binding protein binds to the untranslated regions of prolamine mRNA and to poly(A). Plant Sci. 152: 115–122.Google Scholar
  28. Schultz, J., Milpetz, F., Bork, P. and Ponting, C.P. 1998. SMART, a simple architecture research tool: identification of signalling domains. Proc. Natl. Acad. Sci USA 95: 5857–5864.Google Scholar
  29. Singer, R.H. 1992. The cytoskeleton and mRNA localization. Curr. Opin. Cell Biol. 4: 15–19.Google Scholar
  30. Singer, R.H. 1996. RNA: traffic report. Trends Cell Biol. 6: 486–489. St Johnson, D. 1995. The intracellular localization of messenger RNAs. Cell 81: 161—170.Google Scholar
  31. Takaiwa, F., Ogawa, M. and Okita, T.W. 1999. Rice glutelin. In: R. Casey and P.R. Shewry (Eds.) Seed Proteins, Chapman and Hall, London, pp. 401–426.Google Scholar
  32. Tong, X., Drapkin, R., Yalmanchili, R., Moslalos, G. and Kieff, E. 1995. The Epstein-Barr virus nuclear protein 2 acidic domain forms a complex with a novel cellular coactivator that can interact with TFIIE. Mol. Cell Biol. 15: 4735–4744.Google Scholar
  33. Trendelenburg, G., Hummel, M., Riecken, E.O. and Hanski, C. 1996. Molecular characterization of AKAP149, a novel A kinase anchor protein with a KH domain. Biochem. Biophys. Res. Commun. 5: 313–319.Google Scholar
  34. Wilhelm, J.E. and Vale, R.D. 1993. RNA on the move: the mRNA localization pathway. J. Cell Biol. 123: 269–274.Google Scholar
  35. Yamagata, H. and Tanaka, K. 1986. The site of synthesis and accumulation of rice storage proteins. Plant Cell Physiol. 27: 135–145.Google Scholar
  36. Yisraeli, J.K., Sokol, S. and Melton, D.A. 1990. A two step model for the localization of maternal mRNA in Xenopus oocytes: involvement of microtubules amd microfilaments in the translocation and anchoring of vg1 mRNA. Development 108: 289–298.Google Scholar

Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • R. Sami-Subbu
    • 1
  • Sang-Bong Choi
    • 2
  • Yujia Wu
    • 2
  • Changlin Wang
    • 2
  • Thomas W. Okita
    • 2
  1. 1.National Chemical LaboratoryPuneIndia
  2. 2.Institute of Biological ChemistryWashington State UniversityPullmanUSA

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