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THE PLANT CELL, Vol 6, Issue 5 659-668, Copyright © 1994 by American Society of Plant Biologists
A Protein Binding AT-Rich Sequence in the Soybean Leghemoglobin c3 Promoter Is a General cis Element That Requires Proximal DNA Elements to Stimulate Transcription
N. B. Laursen, K. Larsen, J. Y. Knudsen, H. J. Hoffmann, C. Poulsen, K. A. Marcker and E. O. Jensen
Laboratory of Gene Expression, Department of Molecular Biology, University of Aarhus, Gustav Wieds Vej 10, DK-8000 Aarhus C., Denmark
A nodule nuclear factor, NAT2, interacts with two AT-rich binding sites
(NAT2 BS1 and NAT2 BS2) in the soybean leghemoglobin (lb) c3 promoter. In
transgenic Lotus corniculatus nodules, an oligonucleotide containing NAT2
BS1 activated an inactive -159 lbc3 promoter when placed immediately
upstream of the promoter. The activation was independent of the orientation
of NAT2 BS1 but was dependent on its position in the promoter. The
abilities of different mutated binding sites to activate expression in vivo
were correlated to their respective in vitro affinities for binding NAT2.
This suggested that the interaction between NAT2 and NAT2 BS1 is
responsible for the observed reactivation. Further activation experiments
with the lbc3 and the leaf-specific Nicotiana plumbaginifolia ribulose
bisphosphate carboxylase/oxygenase small subunit (rbcS-8B) promoter
suggested that another specific cis element(s) is required for the function
of NAT2 BS1. Thus, the -102 lbc3 promoter lacking the organ-specific
element (-139 to -102) was not reactivated by the presence of the binding
site, and the rbcS-8B promoter required sequences between -312 and -257 to
be activated by NAT2 BS1. This implies that NAT2 has to work in combination
with other trans-acting factor(s) to increase expression. The finding of
NAT2-like binding activities in different plant organs and the specific
expression of the hybrid NAT2 BS1/-312 rbcS-8B promoter in leaves suggest
that NAT2 is a general activator of transcription.
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