Keratinocyte growth factor regulates expression of the intestinal trefoil factor gene through a cis-regulatory element regulating goblet cell-specific transcription

Intestinal trefoil factor (ITF) is expressed selectively in intestinal goblet cells. Previous studies of the rat ITF gene identified one cis-regulatory element, designated the goblet-cell-response element (GCRE), present in the proximal region of the promoter. To identify additional cis-regulatory elements responsible for goblet-cell-specific expression, a DNA fragment containing 6353 bp of the 5′-flanking region of the mouse ITF gene was cloned and its promoter activity was examined extensively. In human and murine intestinal-derived cell lines (LS174T and CMT-93), the luciferase activities of a 6.3-kb construct were 5- and 2-fold greater than the smaller 1.8-kb construct, respectively. In contrast, the activity in non-intestinal cell lines (HepG2 and HeLa) was 2-4-fold lower than the smaller construct. In the region downstream from the 1.8-kb position, strong luciferase activities in LS174T and HepG2 cells were observed using a 201-bp construct. Interestingly, increased activity was almost completely suppressed in cells transfected with a 391-bp construct. Detailed analyses of this region revealed the existence of a 11-bp positive regulatory element (-181 to -170; ACCTCTTCCTG) and a 9-bp negative regulatory element (-208 to -200; ATTGACAGA) in addition to the GCRE. All three elements were well conserved among human, rat and mouse ITF gene promoters. In addition, a mutant 1.8-kb construct in which the negative regulatory region was deleted yielded the same approximate luciferase activity as a 6.3-kb construct, suggesting binding of a goblet-cell-specific silencer inhibitor (SI) between -6.3 and -1.8 kb. The SI present in goblet cells may block the silencers' binding to the pre-initiation complex and allow increased transcriptional activity driven by specific and non-specific enhancers. High-level expression of the mouse ITF gene specifically in intestinal goblet cells may be achieved through the combined effects of these regulatory elements.

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A silencer inhibitor confers specific expression of intestinal trefoil factor in gobletlike cell lines

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.2001.280.6.g1114 ◽

2001 ◽

Vol 280 (6) ◽

pp. G1114-G1123 ◽

Cited By ~ 15

Author(s):

Dai Iwakiri ◽

Daniel K. Podolsky

Keyword(s):

Cell Lines ◽

Goblet Cell ◽

Regulatory Element ◽

Goblet Cells ◽

Transient Transfection ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Specific Expression ◽

Silencer Element

Intestinal trefoil factor (ITF) is selectively expressed in intestinal goblet cells. Previous studies identified cis-regulatory elements in the proximal promoter of ITF, but these were insufficient to recapitulate the exquisite tissue- and cell-specific expression of native ITF in vivo. Preliminary studies suggested that goblet cell-specific expression of murine ITF requires elements far upstream that include a silencer element that effectively prevents ITF expression in non-goblet cells. Transient transfection studies using native or mutant ITF 5′-flanking sequences identified a region that restores expression in goblet cells. This element, designated goblet cell silencer inhibitor (GCSI) element, enables human and murine goblet cell-like cell lines to override the silencing effect of more proximal elements. The GCSI has no intrinsic enhancer activity and regulates expression only when the silencer element is present. Ligation of GCSI and silencer elements to sucrase-isomaltase conferred goblet cell-specific expression. Goblet cells but not non-goblet cells possess a nuclear protein that binds to the GCSI regulatory element (GCSI binding protein; GCSI-BP). Both transient transfection and gel mobility shift assay studies localize the GCSI and GCSI-BP to −2216 to −2204. We conclude that goblet cell-specific transcription of ITF in vivo depends on a regulatory element designated GCSI.

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Identification of a novel transcriptional regulatory element within the promoter region of the keratinocyte growth factor gene that mediates inducibility to cyclic AMP

Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression ◽

10.1016/s0167-4781(99)00069-x ◽

1999 ◽

Vol 1446 (1-2) ◽

pp. 71-81 ◽

Cited By ~ 8

Author(s):

Jie Zhou ◽

Paul W Finch

Keyword(s):

Growth Factor ◽

Cyclic Amp ◽

Promoter Region ◽

Regulatory Element ◽

Keratinocyte Growth Factor ◽

Factor Gene ◽

Transcriptional Regulatory Element ◽

Growth Factor Gene ◽

Transcriptional Regulatory

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O.01 Regulation of intestinal trefoil factor mRNA by nutritional status and keratinocyte growth factor

Clinical Nutrition ◽

10.1016/s0261-5614(98)80069-1 ◽

1998 ◽

Vol 17 ◽

pp. 1

Author(s):

C.F. Estívariz ◽

L.H. Gu ◽

C.R. Jonas ◽

C.L. Farrell ◽

T.R. Ziegler

Keyword(s):

Growth Factor ◽

Nutritional Status ◽

Keratinocyte Growth Factor ◽

Trefoil Factor ◽

Intestinal Trefoil Factor

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Molecular cloning of mouse intestinal trefoil factor and its expression during goblet cell changes

Biochemical Journal ◽

10.1042/bj3110293 ◽

1995 ◽

Vol 311 (1) ◽

pp. 293-297 ◽

Cited By ~ 36

Author(s):

M Tomita ◽

H Itoh ◽

N Ishikawa ◽

A Higa ◽

H Ide ◽

...

Keyword(s):

Goblet Cell ◽

Goblet Cells ◽

Recovery Phase ◽

Nippostrongylus Brasiliensis ◽

Trefoil Factor ◽

Intestinal Trefoil Factor ◽

Cell Hyperplasia ◽

Reverse Transcription Pcr ◽

Rapid Amplification ◽

Mitf Expression

A cDNA encoding mouse intestinal trefoil factor (mITF) was successfully cloned and sequenced from the small intestine of C57BL/6 mouse by using the combination of reverse transcription-PCR and rapid amplification of cDNA ends methods. The gene was, similar to rat and human ITFs, mainly expressed in the small and large intestine. The mITF expression was up-regulated during the recovery phase after depletion of goblet cells in acetic acid-induced colitis. On the other hand, the expression in the jejunum was not altered, while goblet cell hyperplasia was induced by Nippostrongylus brasiliensis infection. These results suggest that the mITF expression did not simply correlate with the number of goblet cells. The mITF may play an important role in the maintenance and repair of mucosal function of the rectum. Additionally, the mITF in the jejunum may play a role in alteration of the physicochemical nature of goblet cell mucins, thereby affecting the establishment of intestinal helminths.

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