Reference "In vitro binding of nuclear proteins to the barley plastocyanin gene promoter region"

Reference ID

8716

Title

In vitro binding of nuclear proteins to the barley plastocyanin gene promoter region

Source

European journal of biochemistry / FEBS, 1993, vol. 217, pp. 97-104

Authors (2)

Nielsen-P-S

Gausing-K

Abstract

Plastocyanin is a nuclear-encoded chloroplast protein participating in electron
transport during photosynthesis. The plastocyanin gene is expressed in
photosynthetic tissue in a developmentally regulated manner and the expression
is stimulated by light. A genomic clone encoding the plastocyanin precursor was
isolated from a barley (Hordeum vulgare) lambda library using a barley cDNA
clone as a probe and the sequence of a 1.9-kb DNA fragment containing the
plastocyanin gene was determined. TATA and CCAAT boxes are located 34-bp and 68-
bp, respectively, upstream of the transcription start site, the 5'-untranslated
leader is 78 nucleotides long, and the intronless gene has at least two
different polyadenylation sites. DNA sites in the plastocyanin gene that mediate
binding of barley nuclear proteins were mapped by mobility-shift assays with
fragments of the promoter/upstream region. Two of the three specific binding
sites characterised in more detail were found to form complexes with the same
factor in cross-competition experiments. One of these sites, narrowed down to a
17-bp sequence at position -512, contains the consensus binding site for Myb-
like transcription factors. The third specific binding site, located at position
-622, contains the sequence CACGTG which is a high-affinity-binding site for
transcription factors of the basic-region leucine-zipper family.