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THE PLANT CELL, Vol 3, Issue 2 105-113, Copyright © 1991 by American Society of Plant Biologists
Monocot Regulatory Protein Opaque-2 Is Localized in the Nucleus of Maize Endosperm and Transformed Tobacco Plants
M. J. Varagona, R. J. Schmidt and N. V. Raikhel
Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48824-1312
Protein targeting to the nucleus has been studied extensively in animal and
yeast systems; however, nothing is known about nuclear targeting in plants.
The Opaque-2 (O2) gene produces a regulatory protein that is responsible
for inducing transcription of the [alpha]-zein class of storage proteins in
maize kernels. The cloned O2 gene encodes a protein that contains a leucine
zipper DNA binding domain that can interact with zein gene promoters. We
have used immunolocalization to show that the O2 protein is present in
nuclei in the maize endosperm tissues known to produce [alpha]-zeins. In
addition, neither embryo tissue from wild-type kernels nor endosperm from
kernels harboring a null o2 allele contain the O2 protein. Analysis of a
transposable, element-induced o2 allele, o2-m20, revealed that sectors of
endosperm cells contained the nuclear-localized O2 protein, indicating
excision of the transposable element. To study further the nuclear
transport of the O2 protein, we have transformed this gene, under the
control of a constitutive promoter, into tobacco. Plants were shown to have
detectable levels of steady-state O2 mRNA and O2 protein.
Immunolocalization of O2 protein in transformed tobacco plants indicated
that the O2 protein was transported into tobacco nuclei. Therefore, we have
developed a system to study nuclear targeting in plants and have
established that the nuclear transport machinery is similar in monocots and
dicots.
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