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Isolation and characterization of tomato cDNA and genomic clones encoding the ubiquitin gene ubi3

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Abstract

We report here the isolation and nucleotide sequence of tomato cDNA and genomic clones encoding a ubiquitin extension protein homologous to the yeast gene ubi3. Sites similar to upstream activating sites commonly found in the promoters of yeast ribosomal genes were observed in the tomato promoter. The tomato ubi3 promoter also contained elements found in the rbcS promoter from pea. The transcription initiation site was determined to occur 66 bp upstream of the initiating Met. RFLP mapping revealed that the gene was located on chromosome 1, 23 cM from marker TG301. A ubi3 gene-specific probe hybridized to a single 800 nt transcript. Expression was reduced in heat-shocked plants and plants kept in the dark. Expression was highest in young leaves and immature green fruit and lowest in mature leaves and petals. We isolated the original cDNA clone using an antibody prepared against chloroplast polypeptides. Immunological studies did not detect ubiquitin or ubiquitin extension proteins in the chloroplast. However, higher-molecular-weight chloroplast proteins were detected with ubiquitin antisera suggesting that ubiquitin conjugates are transported into the chloroplast.

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Authors and Affiliations

  1. Department of Plant Biology, Carnegie Institute of Washington, 290 Panama Street, 94305, Stanford, CA, USA

    Neil E. Hoffman & Deborah Milkowski

  2. Department of Biology, Queens University, K7L 3N6, Kingston, Ontario, Canada

    Kenton Ko

  3. Biology Department, University of Michigan, 48109, Ann Arbor, MI, USA

    Eran Pichersky

Authors
  1. Neil E. Hoffman
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  2. Kenton Ko
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  3. Deborah Milkowski
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  4. Eran Pichersky
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Hoffman, N.E., Ko, K., Milkowski, D. et al. Isolation and characterization of tomato cDNA and genomic clones encoding the ubiquitin gene ubi3 . Plant Mol Biol 17, 1189–1201 (1991). https://doi.org/10.1007/BF00028735

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  • DOI: https://doi.org/10.1007/BF00028735

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