Cloning of a DNA-binding protein that interacts with the ethylene-responsive enhancer element of the carnation GST1 gene.
Ethylene transcriptionally activates a glutathione S-transferase gene (GST1) at the onset of the senescence program in carnation (Dianthus caryophyllus L.) flower petals. A 126 bp region of the GST1 promoter sequence has been identified as an ethylene-responsive enhancer element (ERE). In this paper, we demonstrate the ability of nuclear proteins from senescing petals to recognize a 22 bp sequence within the ERE (ERE oligonucleotide). Mutation of the ERE oligonucleotide sequence significantly alters the strength of this nuclear protein-DNA association. The wild-type ERE oligonucleotide sequence was used to isolate a cDNA clone encoding a sequence-specific DNA binding protein. Nucleotide sequencing and deduced amino acid sequence analysis of this cDNA predicted a 32 kDa protein which we have designated carnation ethylene-responsive element-binding protein-1 (CEBP-1). The mRNA expression pattern of CEBP-1 suggests that it is not transcriptionally regulated by ethylene. The amino acid sequence homology of CEBP-1 with other plant nucleic acid binding proteins indicates a conserved nucleic acid binding domain. Within this domain are two highly conserved RNA-binding motifs, RNP-1 and RNP-2. An acidic region and a putative nuclear localization signal are also identified.
Maxson
J M
JM
Department of Horticulture, Purdue University, West Lafayette, IN 47907-1165, USA.
Woodson
W R
WR
D000595:Amino Acid Sequence; D001483:Base Sequence; D016922:Cellular Senescence; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005030:Ethylenes; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D005982:Glutathione Transferase / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D059828:Plant Cells; D010937:Plant Growth Regulators; D010940:Plant Proteins; D010944:Plants / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic / Q000187:drug effects
C101503:CEBP-1 protein, Dianthus caryophyllus; D018744:DNA, Plant; D004268:DNA-Binding Proteins; D005030:Ethylenes; D009838:Oligodeoxyribonucleotides; D010937:Plant Growth Regulators; D010940:Plant Proteins; D012333:RNA, Messenger; D018749:RNA, Plant; C036216:ethylene; D005982:Glutathione Transferase