Abstract
Expression of genes (PSPAL) encoding phenylalanine ammonia-lyase from pea (Pisum sativum L.) is regulated in response to various environmental stimuli and during plant development. We examined the cis-regulatory elements in PSPAL1 promoter for organ-specific expression by determining the sequences specifically associated with nuclear proteins on its promoter in each pea organ. In vivo dimethyl sulfate (DMS) footprinting analysis showed putative protein bindings on AC-rich sequences including Box-I in particular in roots and stems in which PSPAL mRNA were highly accumulated. The potential role of the AC-rich element was investigated by fusing PSPAL1 promoter with Box-I deleted to the reporter gene β-glucuronidase (GUS) and transforming the construct into tobacco plants (Nicotiana tabacum). GUS activity controlled under this Box-I deletion promoter was significantly reduced in roots and leaves, whereas drastically increased in stems as compared with the activity of wild-type PSPAL1 promoter. Histochemical GUS staining indicated the activity was elevated in vascular tissues of stem by deleting Box-I. These results suggest that Box-I in PSPAL1 contributes to a positive regulation in root and leaf, but might also function as a negative regulator in stem, especially in xylem tissue.
| Original language | English |
|---|---|
| Pages (from-to) | 355-362 |
| Number of pages | 8 |
| Journal | Plant Physiology and Biochemistry |
| Volume | 39 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2001 |
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Keywords
- Cis-regulatory element
- In vivo footprinting
- Ligation-mediated polymerase chain reaction
- Organ-specific expression
- Phenylalanine ammonia-lyase
- Pisum sativum
ASJC Scopus subject areas
- Plant Science
- Biochemistry
- Biotechnology
Cite this
Contrary operations of Box-I element of pea phenylalanine ammonia-lyase gene 1 promoter for organ-specific expression. / Imura, Y.; Seki, H.; Toyoda, Kazuhiro; Ichinose, Yuki; Shiraishi, T.; Yamada, T.
In: Plant Physiology and Biochemistry, Vol. 39, No. 5, 2001, p. 355-362.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Contrary operations of Box-I element of pea phenylalanine ammonia-lyase gene 1 promoter for organ-specific expression
AU - Imura, Y.
AU - Seki, H.
AU - Toyoda, Kazuhiro
AU - Ichinose, Yuki
AU - Shiraishi, T.
AU - Yamada, T.
PY - 2001
Y1 - 2001
N2 - Expression of genes (PSPAL) encoding phenylalanine ammonia-lyase from pea (Pisum sativum L.) is regulated in response to various environmental stimuli and during plant development. We examined the cis-regulatory elements in PSPAL1 promoter for organ-specific expression by determining the sequences specifically associated with nuclear proteins on its promoter in each pea organ. In vivo dimethyl sulfate (DMS) footprinting analysis showed putative protein bindings on AC-rich sequences including Box-I in particular in roots and stems in which PSPAL mRNA were highly accumulated. The potential role of the AC-rich element was investigated by fusing PSPAL1 promoter with Box-I deleted to the reporter gene β-glucuronidase (GUS) and transforming the construct into tobacco plants (Nicotiana tabacum). GUS activity controlled under this Box-I deletion promoter was significantly reduced in roots and leaves, whereas drastically increased in stems as compared with the activity of wild-type PSPAL1 promoter. Histochemical GUS staining indicated the activity was elevated in vascular tissues of stem by deleting Box-I. These results suggest that Box-I in PSPAL1 contributes to a positive regulation in root and leaf, but might also function as a negative regulator in stem, especially in xylem tissue.
AB - Expression of genes (PSPAL) encoding phenylalanine ammonia-lyase from pea (Pisum sativum L.) is regulated in response to various environmental stimuli and during plant development. We examined the cis-regulatory elements in PSPAL1 promoter for organ-specific expression by determining the sequences specifically associated with nuclear proteins on its promoter in each pea organ. In vivo dimethyl sulfate (DMS) footprinting analysis showed putative protein bindings on AC-rich sequences including Box-I in particular in roots and stems in which PSPAL mRNA were highly accumulated. The potential role of the AC-rich element was investigated by fusing PSPAL1 promoter with Box-I deleted to the reporter gene β-glucuronidase (GUS) and transforming the construct into tobacco plants (Nicotiana tabacum). GUS activity controlled under this Box-I deletion promoter was significantly reduced in roots and leaves, whereas drastically increased in stems as compared with the activity of wild-type PSPAL1 promoter. Histochemical GUS staining indicated the activity was elevated in vascular tissues of stem by deleting Box-I. These results suggest that Box-I in PSPAL1 contributes to a positive regulation in root and leaf, but might also function as a negative regulator in stem, especially in xylem tissue.
KW - Cis-regulatory element
KW - In vivo footprinting
KW - Ligation-mediated polymerase chain reaction
KW - Organ-specific expression
KW - Phenylalanine ammonia-lyase
KW - Pisum sativum
UR - http://www.scopus.com/inward/record.url?scp=0034986595&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034986595&partnerID=8YFLogxK
U2 - 10.1016/S0981-9428(01)01248-7
DO - 10.1016/S0981-9428(01)01248-7
M3 - Article
AN - SCOPUS:0034986595
VL - 39
SP - 355
EP - 362
JO - Plant Physiology and Biochemistry
JF - Plant Physiology and Biochemistry
SN - 0981-9428
IS - 5
ER -