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Nuclease sensitivity of storage-protein genes in isolated nuclei of pea seeds

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Abstract

The chromatin structure of legumin genes in nuclei isolated from leaves and cytyledons of pea (Pisum sativum L.) was investigated. Digestions with micrococcal nuclease (EC 3.1.31.1.) showed that the nucleosomal repeat length of total chromatin (171±25 base pairs) was similar in nuclei from both tissues. The sensitivity to pancreatic deoxyribonuclease (DNase I; EC 3.1.21.1.) of the legumin genes in cotyledon nuclei was greater than that in leaf nuclei; this increase in sensitivity correlated with transcriptional activity of the genes. No DNase-I-hypersensitive sites were detected in these genes in either tissue.

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Abbreviations

bp:

nucleotide base pairs

d.a.f.:

days after flowering

DNase I:

pancreatic deoxyribonuclease

kb:

103 nucleotide base pairs

MNase:

micrococcal nuclease

References

  • Boulter, D. (1981) Biochemistry of storage protein synthesis and deposition in the developing legume seed. Adv. Bot. Res. 9, 1–31

    Google Scholar 

  • Bown, D., Levasseur, M., Croy, R.R.D., Boulter, D., Gatehouse, J.A. (1985) Sequence of a pseudogene in the legumin gene family of pea (Pisum sativum L.). Nucleic Acids Res. 13, 4527–4538

    Google Scholar 

  • Burch, J.B.E., Weintraub, H. (1983) Temporal order of chromatin structural changes associated with activation of the major chicken vitellogenin gene. Cell 33, 65–76

    Google Scholar 

  • Conklin, K.F., Groudine, M. (1984) Chromatin structure and gene expression. In: DNA methylation, pp. 293–351, Razin, A., Cedar, H., Riggs, A.D., eds. Springer, New York

    Google Scholar 

  • Elgin, S.C.R. (1981) DNase I-hypersensitive sites of chromatin. Cell 27, 413–415

    Google Scholar 

  • Elgin, S.C.R. (1984) Anatomy of hypersensitive sites. Nature 309, 213–214

    Google Scholar 

  • Emerson, B.M., Felsenfeld, G. (1984) Specific factor conferring nuclease hypersensitivity at the 5′ end of the chicken adult α-globin gene. Proc. Natl. Acad. Sci. USA 81, 95–99

    Google Scholar 

  • Emerson, B.M., Lewis, C.D., Felsenfeld, G. (1985) Interaction of specific nuclear factors with the nuclease-hypersensitive region of the chicken adult α-globin gene: nature of the binding domain. Cell 41, 21–30

    Google Scholar 

  • Evans, I.M., Croy, R.R.D., Hutchinson, P., Boulter, D., Payne, P.I., Gordon, M.E. (1979) Cell-free synthesis of some storage protein subunits by polyribosomes and RNA isolated from developing seeds of pea (Pisum sativum L.). Planta 144, 455–462

    Google Scholar 

  • Evans, I.M., Gatehouse, J.A., Croy, R.R.D., Boulter, D. (1984) Regulation of the transcription of storage-protein mRNA in nuclei isolated from developing pea (Pisum sativum L.) cotyledons. Planta 160, 559–568

    Google Scholar 

  • Fritton, H.P., Igo-Kemenes, T., Nowock, J., Strech-jurk, U., Theisen, M., Sippel, A.E. (1984) Alternative sets of DNase I-hypersensitive sites characterize the various functional states of the chicken lysozyme gene. Nature 311, 163–165

    Google Scholar 

  • Garel, A., Axel, R. (1976) Selective digestion of transcriptionally active ovalbumin genes from oviduct nuclei. Proc. Natl. Acad. Sci. USA 73, 3966–3970

    Google Scholar 

  • Gatehouse, J.A., Evans, I.M., Bown, D., Croy, R.R.D., Boulter, D. (1982) Control of storage-protein synthesis during seed development in pea (Pisum sativum L.). Biochem. J. 208, 119–127

    Google Scholar 

  • Graham, D.E. (1978) The isolation of high molecular weight DNA from whole organisms or large tissue masses. Anal. Biochem. 85, 609–613

    Google Scholar 

  • Jackson, P.D., Felsenfeld, G. (1985) A method for mapping intranuclear protein-DNA interactions and its application to a nuclease hypersensitive site. Proc. Natl. Acad. Sci. USA 82, 2296–2300

    Google Scholar 

  • Lycett, G.W., Croy, R.R.D., Shirsat A.H., Boulter, D. (1984) The complete nucleotide sequence of a legumin gene from pea (Pisum sativum L.). Nucleic Acids Res. 12, 4493–4506

    Google Scholar 

  • Maniatis, T., Fritsch, E.M., Sambrook, J. (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • McGhee, J.D., Engel, J.D. (1975) Subunit structure of chromatin is the same in plants and animals. Nature 254, 449–450

    Google Scholar 

  • Millerd, A., Spencer, D. (1974) Changes in RNA-synthesising activity and template activity in nuclei from cotyledons of developing pea seeds. Aust. J. Plant. Physiol. 1, 331–341

    Google Scholar 

  • Murray, M.G., Kennard, W.C. (1984) Altered chromatin conformation of the higher plant gene phaseolin. Biochemistry 23, 4225–4232

    Google Scholar 

  • Prentice, D.A., Gurley, L.R. (1983) Nuclease digestibility of chromatin is affected by nuclei isolation procedures. Biochim. Biophys. Acta 740, 134–144

    Google Scholar 

  • Reeves, R. (1984) Transcriptionally active chromatin. Biochim. Biophys. Acta 782, 343–393

    Google Scholar 

  • Sawyer, R.M. (1986) Isolation of a vicilin gene from pea (Pisum sativum L.), and nuclease sensitivity of seed storage protein genes in pea chromatin. Ph.D. thesis, University of Durham

  • Shirsat, A.H. (1984) A gene for legumin: a major storage protein of Pisum sativum L. Ph.D. thesis, University of Durham

  • Smith, D.L. (1973) Nucleic acid, protein and starch synthesis in developing cotyledons of Pisum sativum L. Ann. Bot. 37, 795–804

    Google Scholar 

  • Southern, E.M. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98, 503–517

    Google Scholar 

  • Spiker, S. (1985) Plant chromatin structure. Annu. Rev. Plant Physiol. 36, 235–253

    Google Scholar 

  • Spiker, S., Murray, M.G., Thompson, W.F. (1983) DNase I sensitivity of transcriptionally acitve genes in intact nuclei and isolated chromatin of plants. Proc. Natl. Acad. Sci. USA 80, 815–819

    Google Scholar 

  • Stalder, J., Larsen, A., Engel, J.D., Dolan, M., Groudine, M., Weintraub, H. (1980) Tissue-specific DNA cleavages in the globin chromatin domain introduced by DNase I. Cell 20, 451–460

    Google Scholar 

  • Stalder, J., Seebeck, T., Braun, R. (1979) Accessibility of the ribosomal genes to micro-coccal nuclease in Physarum polycephalum. Biochim. Biophys. Acta 561, 452–463

    Google Scholar 

  • Thomas, P.S., Farquhar, M.N. (1978) Specific measurement of DNA nuclei and nucleic acids using diaminobenzoic acid. Anal. Biochem. 89, 35–44

    Google Scholar 

  • Weintraub, H. (1985) Assembly and propagation of repressed and derepressed chromatin states. Cell 42, 705–711

    Google Scholar 

  • Wu, C. (1980) The 5′ ends of Drosophila heat-shock genes in chromatin are hypersensitive to DNase I. Nature 286, 854–860

    Google Scholar 

  • Wu, C. (1984a) Two protein-binding sites in chromatin implicated in the activation of heat-shock genes. Nature 309, 229–234

    Google Scholar 

  • Wu, C. (1984b) Activating protein factor binds in vitro to upstream control sequences in heat-shock gene chromatin. Nature 311, 81–84

    Google Scholar 

  • Wu, C. (1985) An exonuclease protection assay reveals heatshock element and TATA box DNA-binding proteins in crude nuclear extracts. Nature 317, 84–87

    Google Scholar 

  • Wu, C., Bingham, P.M., Livak, K.J., Holmgren, R., Elgin, S.C.R. (1979) The chromatin structure of specific genes: I. Evidence for higher order domains of defined DNA sequence. Cell 16, 797–806

    Google Scholar 

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Sawyer, R.M., Boulter, D. & Gatehouse, J.A. Nuclease sensitivity of storage-protein genes in isolated nuclei of pea seeds. Planta 171, 254–258 (1987). https://doi.org/10.1007/BF00391102

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  • DOI: https://doi.org/10.1007/BF00391102

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