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J Biol Chem, Vol. 274, Issue 14, 9175-9182, April 2, 1999
From the Laboratorio de Bioquímica y Biología
Molecular, Departamento de Biotecnología-UPM, ETS
Ingenieros Agrónomos, 28040 Madrid, Spain
A barley endosperm cDNA, encoding a
DNA-binding protein of the bZIP class of transcription factors, BLZ2,
has been characterized. The Blz2 mRNA expression is
restricted to the endosperm, where it precedes that of the hordein
genes. BLZ2, expressed in bacteria, binds specifically to the GCN4-like
motif (GLM; 5'-GTGAGTCAT-3') in a 43-base pair oligonucleotide derived
from the promoter region of a Hor-2 gene (B1-hordein). This
oligonucleotide also includes the prolamin box (PB; 5'-TGTAAAG-3').
Binding by BLZ2 is prevented when the GLM is mutated to 5'-GTGctTCtc-3'
but not when mutations affect the PB. The BLZ2 protein is a potent
transcriptional activator in a yeast two-hybrid system where it
dimerizes with BLZ1, a barley bZIP protein encoded by the ubiquitously
expressed Blz1 gene. Transient expression experiments in
co-bombarded developing barley endosperms demonstrate that BLZ2
transactivates transcription from the GLM of the Hor-2 gene
promoter and that this activation is also partially dependent on the
presence of an intact PB. A drastic decrease in GUS activity is
observed in co-bombarded barley endosperms when using as effectors
equimolar mixtures of Blz2 and Blz1 in
antisense constructs. These results strongly implicate the
endosperm-specific BLZ2 protein from barley, either as a homodimer or
as a heterodimer with BLZ1, as an important transcriptional activator
of seed storage protein genes containing the GLM in their promoters.
Barley BLZ2, a Seed-specific bZIP Protein That Interacts with
BLZ1 in Vivo and Activates Transcription from the
GCN4-like motif of B-hordein Promoters in Barley Endosperm
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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