Modified suppression subtractive hybridization identifies an AP2-containing protein involved in metal responses in Physcomitrella patens.
The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up-regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension-cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals.
Cho
Sung Hyun
SH
School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.
Hoang
Quoc
Q
Phee
Jeong Won
JW
Kim
Yun Young
YY
Shin
Hyun Young
HY
Shin
Jeong Sheop
JS
D000595:Amino Acid Sequence; D029681:Arabidopsis Proteins / Q000737:chemistry*; D001483:Base Sequence; D019068:Bryopsida / Q000187:drug effects* / Q000235:genetics; D002999:Clone Cells; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D020224:Expressed Sequence Tags; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D015723:Gene Library; D017343:Genes, Plant; D018398:Homeodomain Proteins / Q000737:chemistry*; D008670:Metals / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA
C499534:APETALA2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D018398:Homeodomain Proteins; D008670:Metals; D009687:Nuclear Proteins; D010940:Plant Proteins; D012333:RNA, Messenger