First published online August 8, 2002; 10.1104/pp.005561
Plant Physiol, September 2002, Vol. 130, pp. 111-119
A Role for the DOF Transcription Factor BPBF in the Regulation of
Gibberellin-Responsive Genes in Barley Aleurone1
Montaña
Mena,2
Francisco Javier
Cejudo,
Ines
Isabel-Lamoneda, and
Pilar
Carbonero*
Laboratorio de Bioquimica y Biologia Molecular, Departamento de
Biotecnologia-Universidad Politécnica de Madrid, Ingenieros
Agronomos, 28040 Madrid, Spain (M.M., I.I.-L., P.C.); and Instituto de
Bioquimica Vegetal y Fotosintesis, Centro de Investigaciones
Cientificas "Isla de la Cartuja," 41092 Sevilla, Spain (F.J.C.)
Functional analyses of a number of hydrolase gene promoters,
induced by gibberellin (GA) in aleurone cells following germination, have identified a GA-responsive complex as a tripartite element containing a pyrimidine box motif 5'-CCTTTT-3'. We describe here that
BPBF, a barley (Hordeum vulgare) transcription
factor of the DOF (DNA-Binding with One Finger) class, previously shown to be an activator of reserve protein encoding genes during
development, also has a role in the control of hydrolase genes
following seed germination. Northern-blot, reverse
transcriptase-polymerase chain reaction, and in situ hybridization
analyses evidenced that the transcripts of the BPBF-encoding gene
(Pbf), besides being present during endosperm
development, are also expressed in aleurone cells of germinated seeds
where they are induced by GA, an effect counteracted by abscisic acid.
Electrophoretic mobility shift assays have shown that the BPBF protein
binds specifically to the pyrimidine box motif in vitro within the
different sequence contexts that naturally occur in the promoters of
genes encoding a cathepsin B-like protease (Al21) and a
low-isoelectric point  -amylase (Amy2/32b), both induced in the aleurone layers in response to GA. In transient expression experiments, BPBF repressed transcription of the
Al21 promoter in GA-treated barley aleurone layers and
reverted the GAMYB-mediated activation of this protease promoter.
1
This work was financed by Ministerio de
Educación y Cultura (Spain; grant nos. PB97-0561 and
PB97-0745), by Ministerio de Ciencia y Tecnología (Spain;
grant no. BMC2000-1483), and by Junta de Andalucia (Spain; grant no.
CVI-0182). M.M. and I.I.-L. were the recipients of a postdoctoral
contract and a PhD scholarship, respectively, from Ministerio de
Educación y Cultura.
2
Present address: Unidad de Biología Vegetal
Facultad de Ciencias del Medio Ambiente, Avenida Carlos III s/n,
45071 Toledo, Spain.
*
Corresponding author; e-mail pcarbonero{at}bit.etsia.upm.es; fax
34-91-3365757.
© 2002 American Society of Plant Physiologists
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