Nucleic Acids Research, 2001, Vol. 29, No. 20 4097-4105
© 2001 Oxford University Press
A novel class of plant-specific zinc-dependent DNA-binding protein that binds to A/T-rich DNA sequences
Graduate School of Agricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Complementary DNA encoding a DNA-binding protein, designated PLATZ1 (plant AT-rich sequence- and zinc-binding protein 1), was isolated from peas. The amino acid sequence of the protein is similar to those of other uncharacterized proteins predicted from the genome sequences of higher plants. However, no paralogous sequences have been found outside the plant kingdom. Multiple alignments among these paralogous proteins show that several cysteine and histidine residues are invariant, suggesting that these proteins are a novel class of zinc-dependent DNA-binding proteins with two distantly located regions, C-x2-H-x11-C-x2-C-x(4–5)-C-x2-C-x(3–7)-H-x2-H and C-x2-C-x(10–11)-C-x3-C. In an electrophoretic mobility shift assay, the zinc chelator 1,10-o-phenanthroline inhibited DNA binding, and two distant zinc-binding regions were required for DNA binding. A protein blot with 65ZnCl2 showed that both regions are required for zinc-binding activity. The PLATZ1 protein non-specifically binds to A/T-rich sequences, including the upstream region of the pea GTPase pra2 and plastocyanin petE genes. Expression of the PLATZ1 repressed those of the reporter constructs containing the coding sequence of luciferase gene driven by the cauliflower mosaic virus (CaMV) 35S90 promoter fused to the tandem repeat of the A/T-rich sequences. These results indicate that PLATZ1 is a novel class of plant-specific zinc-dependent DNA-binding protein responsible for A/T-rich sequence-mediated transcriptional repression.
* To whom correspondence should be addressed. Tel: +81 52 789 4168; Fax: +81 52 789 4296; Email: nagano{at}agr.nagoya-u.ac.jp Present address: Hirofumi Furuhashi, Department of Genetics, The Graduate University for Advanced Studies, Mishima, Shizuoka-ken 411-8540, Japan The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
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