A set of polyclonal and monoclonal antibodies reveals major differences in post-translational modification of the rat HNF1 and vHNF1 homeoproteins

Chouard, Tanguy; Jeannequin, Odile; Rey-Campos, Javier; Yaniv, Moshe; Traincard, François

doi:10.1016/s0300-9084(97)86928-3

Cited by 13 publications

(17 citation statements)

References 24 publications

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“…For immunohistochemistry, sections were sequentially incubated with biotinylated secondary antibodies (Jackson ImmunoResearch), peroxidase-conjugated streptavidin and DAB substrate (both Vector Labs). The following antibodies were used: rabbit anti-Hnf1a-TC284 (1:500) (Chouard, 1997), monoclonal anti-Hnf1b (1:100; clone 3.12) (Chouard et al, 1997), rabbit anti-ezrin (a gift from D. Louvard, Institut Curie, Paris), rabbit anti-chromogranin A+B (1:50; Progen), UEA1-FITC (1:50; Sigma), rabbit anti-lysozyme (1:200; DakoCytomation).…”

Section: Tissue Sample Preparation and Immunohistochemistrymentioning

confidence: 99%

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Hepatocyte nuclear factor 1α and β control terminal differentiation and cell fate commitment in the gut epithelium

et al. 2010

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SUMMARYThe intestinal epithelium is a complex system characterized by massive and continuous cell renewal and differentiation. In this context, cell-type-specific transcription factors are thought to play a crucial role by modulating specific transcription networks and signalling pathways. Hnf1a and b are closely related atypical homeoprotein transcription factors expressed in several epithelia, including the gut. With the use of a conditional inactivation system, we generated mice in which Hnf1b is specifically inactivated in the intestinal epithelium on a wild-type or Hnf1a -/-genetic background. Whereas the inactivation of Hnf1a or Hnf1b alone did not lead to any major intestinal dysfunction, the concomitant inactivation of both genes resulted in a lethal phenotype. Double-mutant animals had defective differentiation and cell fate commitment. The expression levels of markers of all the differentiated cell types, both enterocytes and secretory cells, were affected. In addition, the number of goblet cells was increased, whereas mature Paneth cells were missing. At the molecular level, we show that Hnf1a and b act upstream of the Notch pathway controlling directly the expression of two crucial components: Jag1 and Atoh1. We demonstrate that the double-mutant mice present with a defect in intestinal water absorption and that Hnf1a and b directly control the expression of Slc26a3, a gene whose mutations are associated with chloride diarrhoea in human patients. Our study identifies new direct target genes of the Hnf1 transcription factors and shows that they play crucial roles in both defining cell fate and controlling terminal functions in the gut epithelium. KEY WORDS: Gut epithelium, Hnf1, Commitment, Differentiation, MouseHepatocyte nuclear factor 1a and b control terminal differentiation and cell fate commitment in the gut epithelium

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Section: Tissue Sample Preparation and Immunohistochemistrymentioning

confidence: 99%

“…Chromatin preparation and ChIP were performed as previously described (Gresh et al, 2004). ChIP experiments were performed with rabbit anti-Hnf1a-TC284 (1:500, Chouard, 1997) or rabbit anti-Hnf1b (1:200, clone H-85, Santa Cruz Biotechnology) antibodies. Primers were designed using PrimerExpress 2.0 software.…”

Section: Chromatin Immunoprecipitation (Chip)mentioning

confidence: 99%

Hepatocyte nuclear factor 1α and β control terminal differentiation and cell fate commitment in the gut epithelium

et al. 2010

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“…However, the antiserum we used, a generous gift from Dr. Moshe Yaniv, recognizes both HNF-1␣ and HNF-1␤, although it binds HNF-1␣ with a much greater affinity (62). Therefore, to conclusively determine whether the HNF-1 binding activity detected in HepG2 nuclear extract represents HNF-1␣ or HNF-1␤, we made use of commercially available antisera raised specifically to each isoform of HNF-1.…”

Section: Hnf-1␣ and Hnf-1␤ Selectively Bind The Hnf-1 Site In The G6pasementioning

confidence: 99%

Differential Role of Hepatocyte Nuclear Factor-1 in the Regulation of Glucose-6-phosphatase Catalytic Subunit Gene Transcription by cAMP in Liver- and Kidney-derived Cell Lines

Streeper

Svitek

Goldman

et al. 2000

Journal of Biological Chemistry

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In liver and kidney, the terminal step in gluconeogenesis is catalyzed by glucose-6-phosphatase. To examine the effect of the cAMP signal transduction pathway on transcription of the gene encoding the catalytic subunit of glucose-6-phosphatase (G6Pase), G6Pase-chloramphenicol acetyltransferase (CAT) fusion genes were transiently transfected into either the liver-derived HepG2 or kidney-derived LLC-PK cell line. Co-transfection of an expression vector encoding the catalytic subunit of cAMP-dependent protein kinase (PKA) markedly stimulated G6Pase-CAT fusion gene expression, and mutational analysis of the G6Pase promoter revealed that multiple regions are required for this PKA response in both the HepG2 and LLC-PK cell lines. A sequence in the G6Pase promoter that resembles a cAMP response element is required for the full PKA response in both HepG2 and LLC-PK cells. However, in LLC-PK cells, but not in HepG2 cells, a hepatocyte nuclear factor-1 (HNF-1) binding site was critical for the full induction of G6Pase-CAT expression by PKA. Changing this HNF-1 motif to that for the yeast transcription factor GAL4 reduces the PKA response in LLC-PK cells to the same degree as deleting the HNF-1 site. However, co-transfection of this mutated construct with chimeric proteins comprising the GAL4-DNA binding domain ligated to the coding sequence for HNF-1␣, HNF-1␤, HNF-3, or HNF-4 completely restored the PKA response. Thus, we hypothesize that, in LLC-PK cells, HNF-1 is acting as an accessory factor to enhance PKA signaling through the cAMP response element by altering G6Pase promoter conformation or accessibility rather than specifically affecting some component of the PKA signal transduction pathway.

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“…Then binding was analyzed by acrylamide gel electrophoresis as described above. The HNF-1␣-specific antiserum, designated TC284 (32,33), was a generous gift from Tanguy Chouard and Moshe Yaniv (Institute Pasteur, France), whereas the Egr-1-specific antiserum was purchased from Santa Cruz Biotechnology.…”

mentioning

confidence: 99%

Hepatocyte nuclear factor-1 acts as an accessory factor to enhance the inhibitory action of insulin on mouse glucose-6-phosphatase gene transcription

Streeper

Eaton

Ebert

et al. 1998

Proc. Natl. Acad. Sci. U.S.A.

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Glucose-6-phosphatase catalyzes the terminal step in the gluconeogenic and glycogenolytic pathways. Transcription of the gene encoding the glucose-6-phosphatase catalytic subunit (G6Pase) is stimulated by cAMP and glucocorticoids whereas insulin strongly inhibits both this induction and basal G6Pase gene transcription. Previously, we have demonstrated that the maximum repression of basal G6Pase gene transcription by insulin requires two distinct promoter regions, designated A (from ؊271 to ؊199) and B (from ؊198 to ؊159). Region B contains an insulin response sequence because it can confer an inhibitory effect of insulin on the expression of a heterologous fusion gene. By contrast, region A fails to mediate an insulin response in a heterologous context, and the mutation of region B within an otherwise intact promoter almost completely abolishes the effect of insulin on basal G6Pase gene transcription. Therefore, region A is acting as an accessory element to enhance the effect of insulin, mediated through region B, on G6Pase gene transcription. Such an arrangement is a common feature of cAMP and glucocorticoid-regulated genes but has not been previously described for insulin. A combination of fusion gene and protein-binding analyses revealed that the accessory factor binding region A is hepatocyte nuclear factor-1. Thus, despite the usually antagonistic effects of cAMP͞glucocorticoids and insulin, all three agents are able to use the same factor to enhance their action on gene transcription. The potential role of G6Pase overexpression in the pathophysiology of MODY3 and 5, rare forms of diabetes caused by hepatocyte nuclear factor-1 mutations, is discussed.

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A set of polyclonal and monoclonal antibodies reveals major differences in post-translational modification of the rat HNF1 and vHNF1 homeoproteins

Cited by 13 publications

References 24 publications

Hepatocyte nuclear factor 1α and β control terminal differentiation and cell fate commitment in the gut epithelium

Hepatocyte nuclear factor 1α and β control terminal differentiation and cell fate commitment in the gut epithelium

Differential Role of Hepatocyte Nuclear Factor-1 in the Regulation of Glucose-6-phosphatase Catalytic Subunit Gene Transcription by cAMP in Liver- and Kidney-derived Cell Lines

Hepatocyte nuclear factor-1 acts as an accessory factor to enhance the inhibitory action of insulin on mouse glucose-6-phosphatase gene transcription

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