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PLANT PHYSIOLOGY , Vol 114, Issue 2 643-652, Copyright © 1997 by American Society of Plant Biologists
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GENE REGULATION AND MOLECULAR GENETICS |
Light Regulation of the Abundance of mRNA Encoding a Nucleolin-Like Protein Localized in the Nucleoli of Pea Nuclei
C. G. Tong, S. Reichler, S. Blumenthal, J. Balk, H. L. Hsieh and S. J. Roux
Department of Botany, The University of Texas, Austin, Texas 78713
A cDNA encoding a nucleolar protein was selected from a pea (Pisum sativum)
plumule library, cloned, and sequenced. The translated sequence of the cDNA
has significant percent identity to Xenopus laevis nucleolin (31%), the
alfalfa (Medicago sativa) nucleolin homolog (66%), and the yeast
(Saccharomyces cerevisiae) nucleolin homolog (NSR1) (28%). It also has
sequence patterns in its primary structure that are characteristic of all
nucleolins, including an N-terminal acidic motif, RNA recognition motifs,
and a C-terminal Gly- and Arg-rich domain. By immunoblot analysis, the
polyclonal antibodies used to select the cDNA bind selectively to a 90kD
protein in purified pea nuclei and nucleoli and to an 88-kD protein in
extracts of Escherichia coli expressing the cDNA. In immunolocalization
assays of pea plumule cells, the antibodies stained primarily a region
surrounding the fibrillar center of nucleoli, where animal nucleolins are
typically found. Southern analysis indicated that the pea nucleolin-like
protein is encoded by a single gene, and northern analysis showed that the
labeled cDNA binds to a single band of RNA, approximately the same size as
the cDNA. After irradiation of etiolated pea seedlings by red light, the
mRNA level in plumules decreased during the 1st hour and then increased to
a peak of six times the 0-h level at 12 h. Far-red light reversed this
effect of red light, and the mRNA accumulation from red/far-red light
irradiation was equal to that found in the dark control. This indicates
that phytochrome may regulate the expression of this gene.
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