Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato

Plant Mol Biol. 1993 May;22(2):279-91. doi: 10.1007/BF00014935.

Abstract

We have characterized a genomic clone containing the potato pathogenesis-related genes STH-2 and STH-21. The two genes are found 4 kb apart on the same chromosome and their sequences are highly similar. They present the same transcriptional orientation and are both interrupted by a single intron. A chimaeric gene consisting of 1015 bp of 5'-flanking sequence and part of the first exon of STH-2 fused to the bacterial beta-glucuronidase gene was highly-expressed in tubers of transgenic potato plants after wounding and elicitor treatments. The levels of activity observed in these transgenic plants parallel those observed for the accumulation of STH-2 mRNAs under similar conditions. This indicates that cis-acting elements necessary for the proper activation of the gene are present within 1 kb of 5'-flanking sequences. Functional analysis of 5' deletions of the STH-2/GUS constructs by transient expression in leaf protoplasts revealed the presence of an upstream regulatory sequence between -135 and -52 which contains a TGAC motif, and a possible negative regulatory region between -52 and -28. A factor present in nuclear extracts of wounded potato tubers was found to bind specifically to nucleotides located between -135 to -105, suggesting that this region contains important cis-regulatory elements.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA Mutational Analysis
  • DNA-Binding Proteins / isolation & purification
  • Gene Expression Regulation*
  • Genes, Plant / genetics*
  • Glucuronidase / biosynthesis
  • Molecular Sequence Data
  • Plant Proteins / biosynthesis
  • Plant Proteins / genetics*
  • Plants, Genetically Modified / genetics
  • Recombinant Fusion Proteins / biosynthesis
  • Regulatory Sequences, Nucleic Acid / genetics*
  • Restriction Mapping
  • Sequence Deletion
  • Sequence Homology, Nucleic Acid
  • Solanum tuberosum / genetics*
  • Transformation, Genetic

Substances

  • DNA-Binding Proteins
  • Plant Proteins
  • Recombinant Fusion Proteins
  • pathogenesis-related proteins, plant
  • Glucuronidase

Associated data

  • GENBANK/M29041
  • GENBANK/M29042