The TATA-less, GC-rich porcine dipeptidylpeptidase IV (DPPIV) promoter shows bidirectional activity
Dipeptidylpeptidase IV (DPPIV) is an exopeptidase highly expressed in the brush-border membrane of the small intestine and in the proximal renal tubules. In this paper the 5'-flanking region of the DPPIV gene containing the promoter was sequenced and functionally characterized. The porcine DPPIV promoter lacks a consensus TATA-box, but contains two TATA-like sequences. Evidence for multiple transcription initiation sites was found. Different deletion constructs of the DPPIV 5'-flanking region in front of the CAT gene were analyzed for transient CAT-expression after transfection of the intestinal Caco-2 cell line. These experiments showed that a 89 base pair construct (-91 to -3 relative to the translation initiation site) is sufficient for promoter activity. In the reverse orientation this construct also stimulates transcription with a similar effectivity indicating that the DPPIV promoter has a bidirectional function. The bidirectional function was further demonstrated by the introduction of the -91 to -3 construct into the bidirectional vector system pLUC/CAT-3. In the hepatoma cell line HepG2 two selected deletion constructs (-857 to -3; -282 to -3) were analyzed in the normal orientation using the CAT gene as a reporter gene. The transfection experiments showed that deletion of the region -857 to -282 raised the promoter activity 3-fold. The GC-rich 5'-flanking region was further analyzed and we demonstrate that the DPPIV promoter contains a region with the characteristics of an unmethylated CpG island.
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
1998 |
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| Erschienen: |
1998 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:379 |
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| Enthalten in: |
Biological chemistry - 379(1998), 1 vom: 07. Jan., Seite 75-81 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Qvist, H [VerfasserIn] |
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| Themen: |
Chloramphenicol O-Acetyltransferase |
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| Anmerkungen: |
Date Completed 10.04.1998 Date Revised 18.11.2010 published: Print Citation Status MEDLINE |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM094464804 |
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| 100 | 1 | |a Qvist, H |e verfasserin |4 aut | |
| 245 | 1 | 4 | |a The TATA-less, GC-rich porcine dipeptidylpeptidase IV (DPPIV) promoter shows bidirectional activity |
| 264 | 1 | |c 1998 | |
| 336 | |a Text |b txt |2 rdacontent | ||
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| 500 | |a Date Completed 10.04.1998 | ||
| 500 | |a Date Revised 18.11.2010 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Dipeptidylpeptidase IV (DPPIV) is an exopeptidase highly expressed in the brush-border membrane of the small intestine and in the proximal renal tubules. In this paper the 5'-flanking region of the DPPIV gene containing the promoter was sequenced and functionally characterized. The porcine DPPIV promoter lacks a consensus TATA-box, but contains two TATA-like sequences. Evidence for multiple transcription initiation sites was found. Different deletion constructs of the DPPIV 5'-flanking region in front of the CAT gene were analyzed for transient CAT-expression after transfection of the intestinal Caco-2 cell line. These experiments showed that a 89 base pair construct (-91 to -3 relative to the translation initiation site) is sufficient for promoter activity. In the reverse orientation this construct also stimulates transcription with a similar effectivity indicating that the DPPIV promoter has a bidirectional function. The bidirectional function was further demonstrated by the introduction of the -91 to -3 construct into the bidirectional vector system pLUC/CAT-3. In the hepatoma cell line HepG2 two selected deletion constructs (-857 to -3; -282 to -3) were analyzed in the normal orientation using the CAT gene as a reporter gene. The transfection experiments showed that deletion of the region -857 to -282 raised the promoter activity 3-fold. The GC-rich 5'-flanking region was further analyzed and we demonstrate that the DPPIV promoter contains a region with the characteristics of an unmethylated CpG island | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Chloramphenicol O-Acetyltransferase |2 NLM | |
| 650 | 7 | |a EC 2.3.1.28 |2 NLM | |
| 650 | 7 | |a Dipeptidyl Peptidase 4 |2 NLM | |
| 650 | 7 | |a EC 3.4.14.5 |2 NLM | |
| 700 | 1 | |a Sjöström, H |e verfasserin |4 aut | |
| 700 | 1 | |a Norén, O |e verfasserin |4 aut | |
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