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Plant and Cell Physiology, 1995, Vol. 36, No. 8 1657-1667
© 1995

Interactions of the Glutelin Gt3 5' Flanking Regulatory Regions with Rice Nuclear Proteins

Yuan Zhao and Thomas W. Okita1

Institute of Biological Chemistry, Washington State University Pullman, WA 99164, U.S.A.

1Corresponding author; fax 1-509-335-7643.

Three DNA binding activities, BP-1, BP-2 and BP-3, which interact with the 5' flanking region of the rice glutelin Gt3 gene, were identified by gel retardation assays of rice seed nuclear extracts. The DNA binding activities were seed-specific as identical DNA-protein complexes were not observed when nuclear extracts from leaf tissue or suspension culture cells were analyzed, suggesting that these DNA binding activities are involved in seed-specific expression of the Gt3 gene. The DNA sequences recognized by these DNA binding activities were identified by DNasel footprinting and Bal 31 nuclease mapping analyses. BP-1 recognizes DNA sequences located at –272 bp to –259 bp relative to the transcriptional start site. This DNA segment contains a sequence motif that is conserved among several seed protein genes, implicating that the motif may be a common cis-regulatory element determining seed-specific expression of these genes. BP-2 interacts with sequences located between –861 bp to –838 bp while BP-3 interacts with sequences upstream of –788 bp. The temporal levels of BP-2 binding activity parallel the steady state levels of the Gt3 mRNAs during seed development. Overall, these results and those obtained from in vivo promoter analysis in transgenic plants [Zhao et al. (1994) Plant Mol. Biol. 25: 429] indicate that multiple regulatory elements located at two spatially separated regions of the Gt3 promoter are involved in endosperm-specific and temporal regulation.

(Received July 17, 1995; Accepted October 7, 1995)
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