PLANT PHYSIOLOGY , Vol 108, Issue 1 129-139, Copyright © 1995 by American Society of Plant Biologists
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GENE REGULATION AND MOLECULAR GENETICS |
Characterization of Nuclear Factors for Elicitor-Mediated Activation of the Promoter of the Pea Phenylalanine Ammonia-Lyase Gene 1
H. Kato, M. Wada, K. Muraya, K. Malik, T. Shiraishi, Y. Ichinose and T. Yamada
Laboratory of Plant Pathology and Genetic Engineering, College of Agriculture, Okayama University, Tsushima, Okayama, 700, Japan (H.K., K. Muraya, K. Malik, T.S., Y.I., T.Y.)
The nuclear factors presumably associated with the activation of the gene
encoding phenylalanine ammonia-lyase by a fungal elicitor were
characterized in pea (Pisum sativum L.) epicotyls. The TATA-proximal region
was dissected and putative cis-regulatory elements in the promoter of pea
phenylalanine ammonia-lyase gene 1 were examined by gel-mobility shift and
in vitro footprinting analyses. Specific binding of the nuclear factors to
the promoter-proximal regions of pea phenylalanine ammonia-lyase gene 1
associated with elicitor-mediated activation was detected at a region
containing consensus sequence motifs of boxes 2 and 4 and other AT-rich
sequences. The analyses of DNA fragments containing the deleted promoter
regions suggested that a residue from -183 to -173 (ATTAGTAAGTGAT) was
essential for a maximal activity of forming low-mobility complex (LMC) in
the gel-mobility shift assay, and synthetic oligonucleotides confirmed the
presence of at least one nuclear component associated with the formation of
an active LMC. Competition experiments and treatment with Hoechst 33258
provided direct evidence that the formation of LMC with the promoter
fragments from genes encoding phenylalanine ammonia-lyase and chalcone
synthase in pea contained one or more of the same proteins that recognize
AT-rich sequence motifs for binding. It also suggests that common
high-mobility group-like proteins might be involved in the regulation of
elicitor-inducible genes in pea.
