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Fine structure and function of the osmotin gene promoter

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Abstract

The gene encoding osmotin, a tobacco pathogenesis-related protein, has been shown to be regulated by an array of hormonal and environmental signals. The osmotin promoter fragment −248 to −108 upstream of the transcription start site (fragment A), was sufficient to direct reporter gene expression when fused to a minimal CaMV 35S promoter in transient assays using microprojectile bombardment. This was consistent with previous 5′-deletion analyses of the osmotin promoter which showed that the promoter sequence from −248 to −108 is absolutely required for reporter gene activity. Nuclear protein factors from salt-adapted tobacco cells, ABA-treated unadapted cells, and young cultured tobacco leaves were shown to interact with fragment A by gel mobility-shift assays. DNase I footprinting revealed that three conserved promoter elements in fragment A interact specifically with nuclear factors. These elements are: (1) a cluster of G-box-like sequences (G sequence); (2) an AT-1 box-like sequence, 5′-AATTATTT-TATG-3′ (AT sequence); (3) a sequence highly conserved in ethylene-induced PR gene promoters, 5′-TAAGA/CGCCGCC-3′ (PR sequence). Transient expression assays performed with fragment A deletions fused to GUS indicated that osmotin promoter activity correlated with the presence of these elements. UV cross-linking analysis showed that the protein complex bound to fragment A consisted of at least four individual proteins with approximate molecular masses of 28, 29, 40 and 42 kDa. One component of this protein complex, which was associated with the G sequence, was a 14-3-3 like protein.

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Authors and Affiliations

  1. Center for Plant Environmental Stress Physiology, Purdue University, 1165 Horticulture Building, 47907-1165, West Lafayette, IN, USA

    Dong Liu, Meena L. Narasimhan, Yi Xu, Kashchandra G. Raghothama, Paul M. Hasegawa & Ray A. Bressan

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  1. Dong Liu
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  2. Meena L. Narasimhan
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Liu, D., Narasimhan, M.L., Xu, Y. et al. Fine structure and function of the osmotin gene promoter. Plant Mol Biol 29, 1015–1026 (1995). https://doi.org/10.1007/BF00014974

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