Abstract
Elicitor-induced activation of the potato pathogenesis-related gene PR-10a requires a 30-bp promoter sequence termed the ERE (elicitor response element) that is bound by the nuclear factor PBF-2 (PR-10a binding factor 2). In this study, PBF-2 has been purified to near homogeneity from elicited tubers through a combination of anion-exchange and DNA affinity chromatography. Evidence demonstrates that inactive PBF-2 is stored in the nuclei of fresh tubers and becomes available for binding to the ERE upon elicitation. A protein with an apparent molecular mass of 24 kD (p24) is a DNA binding component of PBF-2. A cDNA encoding p24 has been cloned and encodes a novel protein with a potential transcriptional activation domain that could also act as a single-stranded DNA binding domain. Both PBF-2 and the cDNA-encoded protein bind with high affinity to the single-stranded form of the ERE in a sequence-specific manner. The inverted repeat sequence of the ERE, TGACAnnnnTGTCA, is critical for binding of this factor in vitro and for PR-10a expression in vivo, supporting the role of PBF-2 as a transcriptional regulator.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Antibodies / immunology
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Antibodies / pharmacology
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Base Sequence
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Binding Sites
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Cell Nucleus / chemistry
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Cell Nucleus / drug effects
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Cell Nucleus / metabolism
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Chromatography, Affinity
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Cloning, Molecular
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DNA, Single-Stranded / genetics
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DNA, Single-Stranded / metabolism*
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DNA-Binding Proteins / chemistry
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / isolation & purification
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DNA-Binding Proteins / metabolism*
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Gene Expression Regulation, Plant* / drug effects
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Genes, Plant / genetics*
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Genes, Reporter / genetics
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Models, Genetic
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Molecular Sequence Data
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Molecular Weight
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Mutation
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Nuclear Proteins / chemistry
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Nuclear Proteins / genetics
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Nuclear Proteins / isolation & purification
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Nuclear Proteins / metabolism*
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Peptides / chemistry
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Peptides / genetics
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Plant Growth Regulators / pharmacology
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Plant Proteins*
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Protein Binding / drug effects
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Response Elements / genetics
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Solanum tuberosum / cytology
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Solanum tuberosum / drug effects
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Solanum tuberosum / genetics*
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Substrate Specificity
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Trans-Activators / chemistry
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Trans-Activators / genetics
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Trans-Activators / isolation & purification
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Trans-Activators / metabolism
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Transcriptional Activation / drug effects
Substances
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Antibodies
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DNA, Single-Stranded
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DNA-Binding Proteins
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Nuclear Proteins
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PBF-2 protein, Solanum tuberosum
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Peptides
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Plant Growth Regulators
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Plant Proteins
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Recombinant Proteins
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Trans-Activators
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polyglutamine