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KMID : 0578319910010020145
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Molecules and Cells
1991 Volume.1 No. 2 p.145 ~ p.149
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Interaction of a Nuclear Protein with 5´ Flanking Region of Gamma-Zein Gene from Maize
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So jae-Seong
Kalyanram B. Geetha
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Abstract
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We investigated protein-DNA interactions between the 5´ flanking region of a gamma-zein gene and nuclear proteins isolated from developing maize endosperm. Two distinct DNA regions showed DNA/protein complex formation based on gel retardation assays. Competition experiments suggested that the two sequences interacted with the same protein but with different affinities. Gel retardation assays with various DNA fragments and chemical footprinting analyses delimited the minimum DNA sequences responsible for complex fornation at nucleotides -839 to -810 for the distal sequence and -327 to -235 for the proximal sequence relative to the start of transcription. These two regions share the sequence TTAAAACAAAA, which may serve as the primary recognition sequence. This sequence was protected from digestion in chemical footprinting analyses. From sequence analysis of 5´ flanking 1-kb region of gamma-zein genes from a wild type W64A+, an opaque-2 and a modified opaque-2, the modified opaque-2 mutant which has high levels of gamma-zein contains an extra 16-bp AT-rich sequence at 3´ of the footprinted sequence. The presence of this additional AT-rich sequence appears to increase the affinity of binding of the nuclear protein.
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