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In this study, D A-protein interactions of a one kilobase region proximal to the transcription start site of the zrp2 (Zea mays root-preferential 2) gene promoter was analyzed. The intent of this research was to identify regions of the promoter and D Aprotein interactions, that are responsible for root-preferential transcription. In vitro run-on transcription assays had re ealed dramatically higher levels of transcription in 3-d-old roots than in 5-d-old leaves (Held, 1993). Interactions between regions of the zrp2 promoter and nuclear protein extracts from 3-d-old roots and 5-d-old leaves of maize were analyzed by gel mobility-shift assays. Two D A-protein complexes with each protein extract were observed with the -4 72 to -180 region of the promoter. This region showed 72% nucleotide identity with a 159 bp portion of the first intron from an a-tubulin gene that is expressed preferentially in the roots of maize (Montoliu et al., 1989). AD Aprotein complex with high electrophoretic mobility (Z2BP1) was observed with both root and leaf nuclear protein extracts. Complexes Z2BP2 and Z2BP3 were preferentially formed with root and leaf nuclear protein extracts respecti ely. ubsequently, the proteinbinding region was delimited to a 73 bp (-330 to -258) region, which contained several repeats of two sequence motifs consisting only of As and Ts. The effect of the proteinbinding sequence on transcription of the 1 kbp and the 4.7 kbp promoter fragments (fused to the GU reporter gene) were analyzed by particle bombardment-mediated transient assays in roots and lea es of maize. The transcriptional activities of promoters, with the protein-binding region deleted were compared with their respecti e intact promoters. In roots the deletion resulted in 5-fold-less transcriptional activity with the 1 kbp promoter fragment, while the 4.7 kbp promoter showed no significant decrease. In lea es no ignificant decrease was observed with either 1 kbp or 4.7 kbp promoters. We conclude that this protein-binding region is important in enhancing the le el of transcriptional acti ity in roots. CHAPTER 1: INTRODUCTIO Regulation of Gene Expre ion Plant development and function is critically dependent on the precise control of gene expression. In any one cell at a particular time only a subset of the total genetic information is expressed. The available e idence shows that there are unique and overlapping sets of mRNAs among different organs of a plant (Kamala and Goldberg 198

Protein-binding sequences that may regulate organ-preferential transcription of zrp2, a Zea mays root-preferential gene