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Subcellular localization of MURA and MURB proteins encoded by the maize MuDR transposon

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Abstract

MuDR controls transposition of the Mu transposable element family in Zea maysL. It produces two major transcripts: mudrA and mudrB. mudrA encodes the MURA transposase, but no specific function has been ascribed to mudrB, which lacks strong homology to known genes. Using transient expression assays in onion epidermal cells, we defined three monopartite nuclear localization signals (NLSs) of MURA; each was functionally sufficient for nuclear targeting of MURA:GUS fusion proteins. Interestingly, one NLS (NLS-A3) is produced by the splicing of the third intron. In contrast, there were no clear NLS in MURB, and the major form of MURB aggregated in the cytoplasm. Self-interaction of MURA and of MURB was also shown in a yeast two-hybrid assay. To test whether interactions of MURA and MURB can occur at the level of protein translocation into the nucleus, a cytoplasmically localized MURB:GFP was co-expressed with MURA or with the GUS fusion proteins. Co-expression did not change the localization pattern of either MURA or MURB; MURA and MURB do not detectably interact in a yeast two-hybrid assay. These results suggest that MURA and MURB do not mutually affect their localization, at least in the forms examined here.

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References

  • Bainton, R.J., Kubo, K.M., Feng, J.N. and Craig, N.L. 1993. Tn7 transposition: target DNA recognition is mediated by multiple Tn7-encoded proteins in a purified in vitro system. Cell 72: 931–943.

    Google Scholar 

  • Benito, M.-I. and Walbot, V. 1994. The terminal inverted repeat sequences of MuDR are functionally active promoters in maize cells. Maydica 39: 255–264.

    Google Scholar 

  • Benito, M.-I. and Walbot, V. 1997. Characterization of the maize Mutator transposable element MURA transposase as a DNAbinding protein. Mol. Cell. Biol. 17: 5165–5175.

    Google Scholar 

  • Bennetzen, J.L. 1996. The Mutator transposable element system of maize. Curr. Top. Microbiol. Immunol. 204: 195–229.

    Google Scholar 

  • Bodeau, J.P. and Walbot, V. 1992. Regulated transcription of maize Bronze-2 promoter in electroporated protoplasts requires the C1 and R gene products. Mol. Gen. Genet. 233: 379–387.

    Google Scholar 

  • Boehm, U., Heinlein, M., Behrens, U. and Kunze, R. 1995. One of three nuclear localization signals of maize Activator (Ac) transposase overlaps the DNA-binding domain. Plant J. 7: 441–451.

    Google Scholar 

  • Carrington, J.C., Freed, D.D. and Leinicke, A.J. 1991. Bipartite signal sequence mediates nuclear translocation of the plant potyviral Nla protein. Plant Cell 3: 953–962.

    Google Scholar 

  • Chida, K. and Vogt, P.K. 1992. Nuclear translocation of viral Jun but not of cellular Jun is cell cycle dependent. Proc. Natl. Acad. Sci. USA 89: 4290–4294.

    Google Scholar 

  • Christophe, D., Christophe-Hobertus, C. and Pichon, B. 2000. Nuclear targeting of proteins: how many different signals? Cell. Sign. 12: 337–341.

    Google Scholar 

  • Chu, C.C., Wang, C.C., Sun, C.S., Chen, H., Yin, K.C., Chu, C.Y. and Bi, F.Y. 1975. Establishment of an efficient medium for anther culture of rice through comparative experiments on nitrogen sources. Sci. Sin. 18: 659–668.

    Google Scholar 

  • Cokol, M., Nair, R. and Rost, B. 2000. Finding nuclear localization signals. EMBO Rep. 1: 411–415.

    Google Scholar 

  • Conti, E., Uy, M., Leighton, L., Blobel, G. and Kuriyan, J. 1998. Crystallographic analysis of the recognition of a nuclear localization signal by the nuclear import factor karyopherin ?. Cell 94: 193–204.

    Google Scholar 

  • Dang, C.V. and Lee, W.M.F. 1989. Nuclear and nucleolar targeting sequences of c-erb-A, c-myb, N-myc, p53, HSP70, and HIV tat proteins. J. Biol. Chem. 264: 18019–18023.

    Google Scholar 

  • Dingwall, C., Sharnick, S.V. and Laskey, R.A. 1982. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell 30: 449–458.

    Google Scholar 

  • Donlin, M.J., Lisch, D. and Freeling, M. 1995. Tissue-specific accumulation of MURB, a protein encoded by MuDR, the autonomous regulator of the Mutator transposable element family. Plant Cell 7: 1989–2000.

    Google Scholar 

  • Duttweilier, H.M. 1996. A highly sensitive and non-lethal ?-galactosidase plate assay for yeast. Trends Genet. 12: 340–341.

    Google Scholar 

  • Essers, L., Adolphs, R.H. and Kunze, R. 2000. A highly conserved domain of the maize Activator transposase is involved in dimerization. Plant Cell 12: 211–223.

    Google Scholar 

  • Gietz, D., St. Jean, A., Woods, R.A. and Schiestl, R.H. 1992. Improved method for high efficiency transformation of intact yeast cells. Nucl. Acids Res. 20: 1425–1435.

    Google Scholar 

  • Gordon-Kamm, W.J., Spencer, T.M., Mangano, M.L., Adams, T.R., Daines, R.J., Start, W.G., O'Brien, J.V., Chambers, S.A., Adams, W.R., Willetts, N.G., Rice, T.B., Mackey, C.J., Krueger, R.W., Kausch, A.P. and Lemaux, P.G. 1990. Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell 2: 603–618.

    Google Scholar 

  • Görlich, D. and Mattaj, I.W. 1996. Protein kinesis: nucleocytoplasmic transport. Science 271: 1513–1518.

    Google Scholar 

  • Guiochon-Mantel, A., Loosfelt, H., Lescop, P., Sar, S., Atger, M., Perrot-Applanat, M. and Milgrom, E. 1989. Mechanisms of nuclear localization of the progesterone receptor: evidence for interaction between monomers. Cell 57: 1147–1154.

    Google Scholar 

  • Guo, H., Duong., H., Ma, N. and Lin, C. 1999. The Arabidopsis blue light receptor cryptochrome 2 is a nuclear protein regulated by a blue light-dependent post-transcriptional mechanism. Plant J. 19: 279–287.

    Google Scholar 

  • Hall, M.N., Craik, C. and Hiraoka, Y. 1990. Homeodomain of yeast repressor ?2 contains a nuclear localization signal. Proc. Natl. Acad. Sci. USA 87: 6954–6958.

    Google Scholar 

  • Hershberger, R., Warren, C.A. and Walbot, V. 1991. Mutator activity in maize correlates with the presence and expression of the Mu transposable element Mu9. Proc. Natl. Acad. Sci. USA 88: 10198–10202.

    Google Scholar 

  • Hershberger, R.J., Benito, M.-I., Hardeman, K.J., Warren, C., Chandler, V.L. and Walbot, V. 1995. Characterization of the major transcripts encoded by the regulatory MuDR transposable element of maize. Genetics 140: 1087–1098.

    Google Scholar 

  • Jans, D.A., Xiao, C.-Y. and Lam, M.H.C. 2000. Nuclear targeting signal recognition: a key control point in nuclear transport? BioEssays 22: 532–544.

    Google Scholar 

  • Jefferson, R.A. 1987. Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol. Biol. Rep. 5: 387–405.

    Google Scholar 

  • Kalderon, D., Roberts, B.L., Richardson, W.D. and Smith, A.E. 1984. A short amino acid sequence able to specify nuclear location. Cell 39: 499–509.

    Google Scholar 

  • Kenna, M.A., Brachmann, C.B., Devine, S.E. and Boeke, J.D. 1998. Invading the yeast nucleus: a nuclear localization signal at the C terminus of Ty1 integrase is required for transposition in vivo. Mol. Cell. Biol. 18: 1115–1124.

    Google Scholar 

  • Kim, S.-H., Arnold, D., Lloyd, A. and Roux, S.J. 2001. Antisense expression of an Arabidopsis Ran binding protein renders transgenic roots hypersensitive to auxin and alters auxin-induced root growth and development by arresting mitotic progress. Plant Cell 13: 2619–2630.

    Google Scholar 

  • Kleiner, O., Kircher, S., Harter, K. and Batschauer, A. 1999. Nuclear localization of the Arabidopsis blue light receptor cryptochrome 2. Plant J. 19: 289–296.

    Google Scholar 

  • Krebber, H. and Silver, P.A. 2000. Directing proteins to nucleus by fusion to nuclear localization signal tags. Meth. Enzymol. 327: 283–296.

    Google Scholar 

  • Lanford, R.E. and Butel J.S. 1984. Construction and characterization of an SV40 mutant defective in nuclear transport of T-antigen. Cell 37: 801–813.

    Google Scholar 

  • Le, Q.H., Wright, S., Yu, Z. and Bureau, T. 2000. Transposon diversity in Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 97: 7376–7381.

    Google Scholar 

  • Lisch, D., Girard, L., Donlin, M. and Freeling, M. 1999. Functional analysis of deletion derivatives of the maize transposon MuDR delineates roles for the MURA and MURB proteins. Genetics 151: 331–341.

    Google Scholar 

  • Malim, M.H., Böhnlein, S., Hauber, J. and Cullen, B.R. 1989. Functional dissection of the HIV-1 Rev trans-activator: derivation of a trans-dominant repressor of Rev function. Cell 58: 205–214.

    Google Scholar 

  • Maniatis, T., Fritsch, E.F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Mizuuchi, M., Baker, T.A. and Mizuuchi, K. 1995. Assembly of phage Mu transpososomes: cooperative transitions assisted by protein and DNA scaffolds. Cell 83: 375–385.

    Google Scholar 

  • Moreland, R.B., Langevin, G.L., Singer, R.H., Garcea, R.L. and Hereford, L.M. 1987. Amino acid sequences that determine the nuclear localization of yeast histone 2B. Mol. Cell. Biol. 7: 4048–4057.

    Google Scholar 

  • O'Neil, K.T., Hoess, R.H. and DeGrado, W.F. 1990. Design of DNA-binding peptides based on the leucine zipper motif. Science 249: 774–778.

    Google Scholar 

  • Picard, D. and Yamamoto, K.R. 1987. Two signals mediate hormone-dependent nuclear localization of the glucocorticoid receptor. EMBO J. 6: 3333–3340.

    Google Scholar 

  • Poulat, F., Girard, F., Chevron, M.-P., Gozé, A., Rebillard, X., Calas, B., Lamb, N. and Berta, P. 1995. Nuclear localization of the testis determining gene product SRY. J. Cell Biol. 128: 737–748.

    Google Scholar 

  • Raizada, M.N. 2000. Altering the Mutator transposon family to study developmental switches and to facilitate functional genomics. Ph.D. dissertation, Stanford University, Stanford, CA.

    Google Scholar 

  • Raizada, M.N. and Walbot, V. 2000. The late developmental pattern of Mu transposon excision is conferred by a cauliflower mosaic virus 35S-driven MURA cDNA in transgenic maize. Plant Cell 12: 5–21.

    Google Scholar 

  • Röttgers, K., Krohn, N.M., Lichota, J., Stemmer, C., Merkle, T. and Grasser, K.D. 2000. DNA-interactions and nuclear localisation of the chromosomal HMG domain protein SSRP1 from maize. Plant J. 23: 395–405.

    Google Scholar 

  • Rudenko, G.N. and Walbot, V. 2001. Expression and posttranscriptional regulation of maize transposable element MuDR and its derivatives. Plant Cell 13: 553–570.

    Google Scholar 

  • Sakamoto, K. and Nagatani, A. 1996. Nuclear localization activity of phytochrome B. Plant J 10: 859–868.

    Google Scholar 

  • Sambrook, J., Fritsch, E. F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • Sato, T., Hanada, M., Bodrug, S., Irie, S., Iwama, N., Boise, L.H., Thompson, C.B., Golemis, E., Fong, L., Wang, H.-G. and Reed, J.C. 1994. Interactions among members of the Bcl-2 protein family analyzed with a yeast two-hybrid system. Proc. Natl. Acad. Sci. USA 91: 9238–9242.

    Google Scholar 

  • Shieh, M.W., Wessler, S.R. and Raikhel, N.V. 1993. Nuclear targeting of the maize R protein requires two nuclear localization sequences. Plant Physiol. 101: 353–361.

    Google Scholar 

  • Singer, T., Yordan, C. and Martienssen, R.A. 2001. Robertson's Mutator transposons in A. thaliana are regulated by the chromatinremodeling gene Decrease in DNA Methylation (DDM). Genes Dev. 15: 591–602.

    Google Scholar 

  • Sock, E., Enderich, J., Rosenfeld, M.G. and Wegner, M. 1996. Identification of the nuclear localization signal of the POU domain protein Tst-1/Oct6. J. Biol. Chem. 271: 17512–17518.

    Google Scholar 

  • Stacey, M.G., Hicks, S.N. and von Arnim, A.G. 1999. Discrete domains mediate the light-responsive nuclear and cytoplasmic localization of Arabidopsis COP1. Plant Cell 11: 349–363.

    Google Scholar 

  • Summers, M.F., South, T.L., Kim, B. and Hare, D.R. 1990. Highresolution structure of an HIV zinc fingerlike domain via a new NMR-based distance geometry approach. Biochemistry 29: 329–340.

    Google Scholar 

  • Tapscott, S.J., Davis, R.L., Thayer, M.J., Cheng, P.-F., Weintraub, H. and Lassar, A.B. 1988. MyoD1: a nuclear phosphoprotein requiring a Myc homology region to convert fibroblasts to myoblasts. Science 242: 405–411.

    Google Scholar 

  • Varagona, M.J. and Raikhel, N.V. 1994. The basic domain in the bZIP regulatory protein Opaque2 serves two independent functions: DNA binding and nuclear localization. Plant J. 5: 207–214.

    Google Scholar 

  • von Arnim, A.G. and Deng, X.-W. 1994. Light inactivation of Arabidopsis photomorphogenic repressor COP1 involves a cellspecific regulation of its nucleocytoplasmic partitioning. Cell 79: 1035–1045.

    Google Scholar 

  • Walbot, V. and Rudenko, G.N. 2002. MuDR/Mu transposable elements of maize. In: N.L. Craig, R. Craigie, M. Gellert and A. Lambowitz (Eds) Mobile DNA II, American Society for Microbiology, Washington, DC, pp. 533–564.

    Google Scholar 

  • Yu, Z., Wright, S.I. and Bureau, T.E. 2000. Mutator-like elements in Arabidopsis thaliana: structure, diversity and evolution. Genetics 156: 2019–2031.

    Google Scholar 

  • Zhao, L.J. and Padmanabhan, R. 1988. Nuclear transport of adenovirus DNA polymerase is facilitated by interaction with preterminal protein. Cell 55: 1005–1015.

    Google Scholar 

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  1. Department of Biological Sciences, Stanford University, 385 Serra Mall, Stanford, CA , 94305-5020, USA

    Akemi Ono, Soo-Hwan Kim & Virginia Walbot

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  1. Akemi Ono
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  2. Soo-Hwan Kim
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Ono, A., Kim, SH. & Walbot, V. Subcellular localization of MURA and MURB proteins encoded by the maize MuDR transposon. Plant Mol Biol 50, 599–611 (2002). https://doi.org/10.1023/A:1019970206057

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