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Article: Functional identification of an intronic promoter of the human glucose-dependent insulinotropic polypeptide gene

TitleFunctional identification of an intronic promoter of the human glucose-dependent insulinotropic polypeptide gene
Authors
KeywordsGene regulation
GIP
Intestine
Pancreas
Progesterone receptor
Transcription factor II D
Issue Date2010
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/gene
Citation
Gene, 2010, v. 463 n. 1-2, p. 29-40 How to Cite?
AbstractGlucose-dependent insulinotropic polypeptide (GIP), a physiological incretin and enterogastrone, plays a vital role in regulating glucose-dependent insulin release from the pancreas and gastric acid secretion from the stomach. By using a transgenic mouse approach, we previously reported that the distal 1.2. kb promoter region of the human GIP (hGIP) gene (-2545/-346, relative to the ATG) was able to target the transgene expression in the stomach but not in the small intestine where the majority of GIP-producing cells are located. In the present study, in order to identify the cis-acting element(s) that is/are required for intestinal expression, a 1.6. kb (-1580/-) DNA fragment within the first intron of the hGIP gene was isolated and characterized in three GIP-expressing cell lines including HuTu80 (duodenal cells), PANC-1 (pancreatic ductal cells) and Hs746T (stomach cells). By 5' and 3' deletion analysis, a proximal promoter element was confined within the nucleotides -102/-1. This promoter element, functions in an orientation-dependent manner, was able to drive 15.1 and 18.3 fold increases in promoter activities in HuTu80 and PANC-1 cells, respectively. Site-directed mutation analysis indicated that the region -54/-23 was essential for promoter function while the region -22/-1 might possess opposite effects in HuTu80 and PANC-1 cells. In competitive and antibody supershift assays, interactions of the progesterone receptor (PR) and some unknown protein factors from HuTu80 and PANC-1 with the motif(s) at -54/-23 were evident. Consistent with this finding, we demonstrated the transcriptional regulation of the hGIP promoter by progesterone via the PR-B isoform and that progesterone treatment in both HuTu80 and PANC-1 cells resulted in an increase in hGIP transcript level. In addition, a sequence motif (ACATGT) residing -48/-43 was found to be responsible for the binding of potential TFII regulator(s). Taken together, our results suggest that the proximal intronic sequences contain essential cis-acting elements for the cell-specific expression of the hGIP gene. © 2010 Elsevier B.V.
Persistent Identifierhttp://hdl.handle.net/10722/91644
ISSN
2015 Impact Factor: 2.319
2015 SCImago Journal Rankings: 1.043
ISI Accession Number ID
Funding AgencyGrant Number
CRCG20051159086
Funding Information:

The authors would like to thank Dr. Peter C.K. Leung for providing human PR-A and PR-B expression vectors, and Elisa Lau for the preparation of the manuscript. This work was supported by CRCG 20051159086 (to BKCC).

References

 

DC FieldValueLanguage
dc.contributor.authorHoo, RLCen_HK
dc.contributor.authorChu, JYSen_HK
dc.contributor.authorYuan, Yen_HK
dc.contributor.authorYeung, CMen_HK
dc.contributor.authorChan, KYYen_HK
dc.contributor.authorChow, BKCen_HK
dc.date.accessioned2010-09-17T10:22:41Z-
dc.date.available2010-09-17T10:22:41Z-
dc.date.issued2010en_HK
dc.identifier.citationGene, 2010, v. 463 n. 1-2, p. 29-40en_HK
dc.identifier.issn0378-1119en_HK
dc.identifier.urihttp://hdl.handle.net/10722/91644-
dc.description.abstractGlucose-dependent insulinotropic polypeptide (GIP), a physiological incretin and enterogastrone, plays a vital role in regulating glucose-dependent insulin release from the pancreas and gastric acid secretion from the stomach. By using a transgenic mouse approach, we previously reported that the distal 1.2. kb promoter region of the human GIP (hGIP) gene (-2545/-346, relative to the ATG) was able to target the transgene expression in the stomach but not in the small intestine where the majority of GIP-producing cells are located. In the present study, in order to identify the cis-acting element(s) that is/are required for intestinal expression, a 1.6. kb (-1580/-) DNA fragment within the first intron of the hGIP gene was isolated and characterized in three GIP-expressing cell lines including HuTu80 (duodenal cells), PANC-1 (pancreatic ductal cells) and Hs746T (stomach cells). By 5' and 3' deletion analysis, a proximal promoter element was confined within the nucleotides -102/-1. This promoter element, functions in an orientation-dependent manner, was able to drive 15.1 and 18.3 fold increases in promoter activities in HuTu80 and PANC-1 cells, respectively. Site-directed mutation analysis indicated that the region -54/-23 was essential for promoter function while the region -22/-1 might possess opposite effects in HuTu80 and PANC-1 cells. In competitive and antibody supershift assays, interactions of the progesterone receptor (PR) and some unknown protein factors from HuTu80 and PANC-1 with the motif(s) at -54/-23 were evident. Consistent with this finding, we demonstrated the transcriptional regulation of the hGIP promoter by progesterone via the PR-B isoform and that progesterone treatment in both HuTu80 and PANC-1 cells resulted in an increase in hGIP transcript level. In addition, a sequence motif (ACATGT) residing -48/-43 was found to be responsible for the binding of potential TFII regulator(s). Taken together, our results suggest that the proximal intronic sequences contain essential cis-acting elements for the cell-specific expression of the hGIP gene. © 2010 Elsevier B.V.en_HK
dc.languageengen_HK
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/geneen_HK
dc.relation.ispartofGeneen_HK
dc.subjectGene regulationen_HK
dc.subjectGIPen_HK
dc.subjectIntestineen_HK
dc.subjectPancreasen_HK
dc.subjectProgesterone receptoren_HK
dc.subjectTranscription factor II Den_HK
dc.subject.meshGIP-
dc.subject.meshIntestine-
dc.subject.meshPancreas-
dc.subject.meshGene regulation-
dc.subject.meshProgesterone receptor-
dc.titleFunctional identification of an intronic promoter of the human glucose-dependent insulinotropic polypeptide geneen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0378-1119&volume=463&issue=1-2&spage=29&epage=40&date=2010&atitle=Functional+identification+of+an+intronic+promoter+of+the+human+glucose-dependent+insulinotropic+polypeptide+gene-
dc.identifier.emailHoo, RLC: rubyhoo@hkucc.hku.hken_HK
dc.identifier.emailChu, JYS: hitan@graduate.hku.hken_HK
dc.identifier.emailChow, BKC: bkcc@hku.hken_HK
dc.identifier.authorityHoo, RLC=rp01334en_HK
dc.identifier.authorityChu, JYS=rp00684en_HK
dc.identifier.authorityChow, BKC=rp00681en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.gene.2010.04.017en_HK
dc.identifier.pmid20452407en_HK
dc.identifier.scopuseid_2-s2.0-77954657008en_HK
dc.identifier.hkuros175277-
dc.identifier.hkuros226350-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77954657008&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume463en_HK
dc.identifier.issue1-2en_HK
dc.identifier.spage29en_HK
dc.identifier.epage40en_HK
dc.identifier.isiWOS:000280751600005-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridHoo, RLC=6602369766en_HK
dc.identifier.scopusauthoridChu, JYS=34975209300en_HK
dc.identifier.scopusauthoridYuan, Y=55474939100en_HK
dc.identifier.scopusauthoridYeung, CM=26531966700en_HK
dc.identifier.scopusauthoridChan, KYY=25225022200en_HK
dc.identifier.scopusauthoridChow, BKC=7102826193en_HK
dc.identifier.citeulike7147672-

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