First published online June 5, 2003; 10.1104/pp.103.022574
Plant Physiology 132:1475-1488 (2003)
© 2003 American Society of Plant Biologists
DEVELOPMENT AND HORMONE ACTION
Molecular and Biochemical Characterization of VR-EILs Encoding Mung Bean ETHYLENE INSENSITIVE3-LIKE Proteins1
Jae-Hoon Lee and
Woo Taek Kim*
Department of Biology, College of Science, Yonsei University, Seoul
120–749, Korea
ETHYLENE INSENSITIVE3 (EIN3) is a transcription factor involved in the
ethylene signal transduction pathway in Arabidopsis. Two full-length cDNA
clones, pVR-EIL1 and pVR-EIL2, encoding EIN3-LIKE proteins were isolated by
reverse transcriptase-polymerase chain reaction and by screening the cDNA
library of mung bean (Vigna radiata) hypocotyls. VR-EIL1 and VR-EIL2
share 70% identity and display varying degrees of sequence conservation
(39%–65%) with previously isolated EIN3 homologs from Arabidopsis,
tobacco (Nicotiana tabacum) and tomato (Lycopersicon
esculentum) plants. Gel retardation assay revealed that both VR-EILs were
able to interact specifically with optimal binding sequence-1, the recently
identified optimal binding sequence for tobacco TEIL, with the binding of
VR-EIL2 being more efficient than that of VR-EIL1. Transient expression
analysis using a VR-EIL::smGFP fusion gene in onion (Allium
cepa) epidermal cells indicated that the VR-EIL proteins were effectively
targeted to the nucleus. The fusion protein of VR-EIL2 with GAL4 DNA-binding
domain strongly activated transcription of a reporter gene in yeast cells, and
an essential domain for transcription-stimulating activity was localized to
the amino-terminal acidic region that consists of 50 amino acid residues. In
contrast with what has been previously found in EIN3- and
TEIL-overexpressing Arabidopsis plants, transgenic tobacco seedlings
expressing the VR-EIL genes under the control of cauliflower mosaic
virus 35S promoter did not exhibit a constitutive triple response. Instead,
they displayed a markedly enhanced proliferation of root hairs, one of the
typical ethylene response phenotypes, and increased sensitivity to exogenous
ethylene. In addition, the pathogenesis-related (PR) genes encoding
 -1,3-glucanase, osmotin, and PR1 were constitutively expressed in
35S::VR-EIL lines without added ethylene, and were hyperinduced in
response to ethylene treatment. These results indicate that VR-EILs are
functional in tobacco cells, thereby effectively transactivating the
GCC-box-containing PR genes and enhancing sensitivity to ethylene. The
possible physiological role of VR-EILs is discussed in the light of the
suggestion that they are active components of the ethylene-signaling pathway
and their heterologous expressions constitutively turn on a subset of ethylene
responses in tobacco plants.
Article, publication date, and citation information can be found at
www.plantphysiol.org/cgi/doi/10.1104/pp.103.022574.
1 This work was supported by the Plant Diversity Research Center (21st
Century Frontier Research Program of Ministry of Science and Technology
project no. PF 003105–01) and by Korea Science and Engineering
Foundation (Plant Metabolism Research Center, Kyung Hee University, to
W.T.K.).
*
Corresponding author; e-mail
wtkim{at}yonsei.ac.kr;
fax 82–2–312–5657.
Received February 21, 2003;
returned for revision April 2, 2003;
accepted April 2, 2003.
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