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J Behrens

Publications and source records attributed to J Behrens.

At least 37 records · Page 2Linked to original sources
PubMed1998

Functional interaction of an axin homolog, conductin, with beta-catenin, APC, and GSK3beta.

Control of stability of beta-catenin is central in the wnt signaling pathway. Here, the protein conductin was found to form a complex with both beta-catenin and the tumor suppressor gene product adenomatous polyposis coli (APC). Conductin induced beta-catenin degradation, whereas mutants of conductin that were deficient in complex formation stabilized beta-catenin. Fragments of APC that contained a conductin-binding domain also blocked beta-catenin degradation. Thus, conductin is a component of the multiprotein complex that directs beta-catenin to degradation and is located downstream of APC. In Xenopus embryos, conductin interfered with wnt-induced axis formation.

Adenomatous Polyposis Coli Protein↗
PubMed1998

RB and c-Myc activate expression of the E-cadherin gene in epithelial cells through interaction with transcription factor AP-2.

E-cadherin plays a pivotal role in the biogenesis of the first epithelium during development, and its down-regulation is associated with metastasis of carcinomas. We recently reported that inactivation of RB family proteins by simian virus 40 large T antigen (LT) in MDCK epithelial cells results in a mesenchymal conversion associated with invasiveness and a down-regulation of c-Myc. Reexpression of RB or c-Myc in such cells allows the reexpression of epithelial markers including E-cadherin. Here we show that both RB and c-Myc specifically activate transcription of the E-cadherin promoter in epithelial cells but not in NIH 3T3 mesenchymal cells. This transcriptional activity is mediated in both cases by the transcription factor AP-2. In vitro AP-2 and RB interaction involves the N-terminal domain of AP-2 and the oncoprotein binding domain and C-terminal domain of RB. In vivo physical interaction between RB and AP-2 was demonstrated in MDCK and HaCat cells. In LT-transformed MDCK cells, LT, RB, and AP-2 were all coimmunoprecipitated by each of the corresponding antibodies, and a mutation of the RB binding domain of the oncoprotein inhibited its binding to both RB and AP-2. Taken together, our results suggest that there is a tripartite complex between LT, RB, and AP-2 and that the physical and functional interactions between LT and AP-2 are mediated by RB. Moreover, they define RB and c-Myc as coactivators of AP-2 in epithelial cells and shed new light on the significance of the LT-RB complex, linking it to the dedifferentiation processes occurring during tumor progression. These data confirm the important role for RB and c-Myc in the maintenance of the epithelial phenotype and reveal a novel mechanism of gene activation by c-Myc.

3T3 Cells↗
PubMed1998

Iron in prenatal multivitamin/multimineral supplements. Bioavailability.

OBJECTIVE: To determine the effect of the differences in the content of essential and nonessential ingredients and of a standardized meal on iron bioavailability from four popular prenatal multivitamin/multimineral supplements. STUDY DESIGN: Iron absorption during an eight-hour period following ingestion of a multivitamin/multimineral formulation, both fasting and postprandial, after a standardized meal, was measured in five groups of 20 pregnant women (24-32 weeks of gestation) and statistically compared. The prenatal formulations were Natalins Rx, Prenatal 1/1, Stuartnatal Plus and Prenate 90. One-A-Day (without iron) was included in the study for the control group. RESULTS: The descending order of absorption, both fasting and postprandial, were Prenate 90, Stuartnatal Plus, Prenatal 1/1 and Natalins Rx. Total iron absorption for each formulation group during the postprandial phase was higher than during the fasting phase. However, the net amount of iron absorption (in both the fasting and postprandial phases) from three of the four formulations (Stuartnatal Plus, Prenatal 1/1 and Prenate 90) provided the minimal 3.0 mg of supplemental iron per day recommended by the National Academy of Science. CONCLUSION: The observed differences in iron absorption between prenatal supplements apparently reflect the effects of the various combinations of vitamins, essential minerals and additives. The absolute amount of elemental iron contained in a prenatal multivitamin/ multimineral formulation does not ensure availability.

Absorption↗
PubMed1997

Analysis of the E-cadherin and P-cadherin promoters in murine keratinocyte cell lines from different stages of mouse skin carcinogenesis.

We previously isolated the 5' upstream sequences of the mouse P-cadherin gene, in which putative binding sites for several transcription factors were identified between nt-101 and +30. In the study reported here, the promoter activity of the postulated 5' cis-acting sequences of the P-cadherin promoter, and the activity of the proximal E-cadherin promoter were investigated in several murine keratinocyte cell lines showing different levels of P- and E-cadherin expression as well as different morphology and tumorigenic behavior. Cell-type specificity and optimal activity of P-cadherin expression in murine keratinocytes was conferred by 5' sequences located between nt -200 and +30, and the GC-rich region (nt -101 to +80) and a CCAAT box element (nt -65) had a major regulatory role. The cell-type specificity of the E-cadherin promoter, on the other hand, was mediated by a combination of positive regulatory elements, a GC-rich region (nt -58 to -24), and a CCAAT box (nt -65) and repressor elements inside the E-pal sequence. Interestingly, the maximum repressor effect of the E-pal element was observed in non-expressing undifferentiated spindle cells. In vitro binding studies indicated that the GC-rich region of the P-cadherin promoter was mainly recognized by Sp1-related nuclear factors, whereas both AP2- and Sp1-related factors were involved in the interaction of the GC-rich region of the E-cadherin promoter. Common factors (probably related to the CP1 family) seemed also to be involved in the recognition of the CCAAT-box element of both the E- and P-cadherin promoters, but additional specific factors participated in the interaction with the CCAAT box of the E-cadherin promoter. Our studies also support the hypothesis that loss or modification of some of the regulatory factors occurs during mouse skin tumor progression.

Animals↗
PubMed1997

Kikuchi disease (histiocytic necrotizing lymphadenitis) in association with HTLV1.

We report a 25-year-old male with fever, generalized lymphadenopathy and an erythematosus maculo-papular eruption affecting face and upper body. A diagnosis of Kikuchi disease (necrotizing histiocytic lymphadenitis) was made on lymph node histology. The patient was found to be positive for the human T-cell leukaemia lymphoma virus (HTLV1). Kikuchi disease has been reported in association with infections such as Epstein-Barr virus (EBV), herpes (HHV6) and toxoplasmosis, but to our knowledge this is the first case of Kikuchi disease associated with HTLV1.

Adult↗
PubMed1996

Interaction between Gab1 and the c-Met receptor tyrosine kinase is responsible for epithelial morphogenesis.

The proteins Gab1 and the related DOS (for 'daughter of sevenless') each bind to substrates of tyrosine kinases like Grb2 or Corkscrew, and act in signalling pathways downstream of tyrosine kinase receptors. Here we show that Gab1 interacts directly with the c-met-encoded receptor tyrosine kinase but not with a number of other tyrosine kinases from different subfamilies. A newly identified proline-rich domain of Gab1 is responsible for the binding of this protein to the tyrosine-phosphorylated bidentate docking site in c-Met. Expression of Gab1 in epithelial cells is sufficient to induce the c-Met-specific activities, including branching morphogenesis. Thus we have discovered a new phosphotyrosine interaction domain in Gab1 and shown that Gab1 is the substrate of the c-Met receptor tyrosine kinase that mediates epithelial morphogenesis.

Adaptor Proteins, Signal Transducing↗
PubMed1996

Functional interaction of beta-catenin with the transcription factor LEF-1.

The cytoplasmic proteins beta-catenin of vertebrates and armadillo of Drosophila have two functions: they link the cadherin cell-adhesion molecules to the cytoskeleton, and they participate in the wnt/wingless signal pathway. Here we show, in a yeast two-hybrid screen, that the architectural transcription factor LEF-1 (for lymphoid enhancer-binding factor) interacts with beta-catenin. In mammalian cells, coexpressed LEF-1 and beta-catenin form a complex that is localized to the nucleus and can be detected by immunoprecipitation. Moreover, LEF-1 and beta-catenin form a ternary complex with DNA that splays an altered DNA bend. Microinjection of LEF-1 into XenoPus embryos induces axis duplication, which is augmented by interaction with beta-catenin. Thus beta-catenin regulates gene expression by direct interaction with transcription factors such as LEF-1, providing a molecular mechanism for the transmission of signals, from cell-adhesion components or wnt protein to the nucleus.

Animals↗
PubMed1996

Mechanisms identified in the transcriptional control of epithelial gene expression.

Epithelium-specific gene expression is fundamental in both embryogenesis and the maintenance of adult tissues, and impairment of epithelial characteristics contributes to diseases such as cancer. We have here analyzed the 5'-region of the epithelial (E-) cadherin gene in order to understand mechanisms of epithelium-specific transcription and loss of expression during epithelial-mesenchymal transitions. The regulatory region of the mouse epithelial cadherin gene is composed of a promoter (from position -94 to the transcription start site) and a 150-base pair enhancer located in the first intron. The 5'-promoter consists of positive regulatory elements (a CCAAT-box and two AP-2 binding sites in a GC-rich region) and the palindromic element E-Pal that activates and represses transcription in epithelial and mesenchymal cells, respectively. The enhancer of the first intron stimulates the activity of heterologous promoters exclusively in epithelial cells. This epithelium-specific enhancer consists of three elements (E I to E III; E II and E III bind AP-2) that are necessary and sufficient for activity. We thus propose two regulatory mechanisms by which epithelial specificity of epithelial cadherin expression is determined: suppression of promoter activity in mesenchymal cells by E-Pal and enhancement of activity in epithelial cells by both E-Pal and the epithelium-specific enhancer.

Animals↗
PubMed1995

Progression of carcinoma cells is associated with alterations in chromatin structure and factor binding at the E-cadherin promoter in vivo.

E-cadherin has been identified as a tumor (invasion) suppressor gene, which is mutated in 50% of diffuse-type human gastric carcinomas. In other carcinomas, the expression of E-cadherin is down-regulated in the poorly differentiated cells such as from breast, bladder, lung and colon. We have here examined the in vivo properties of the genomic E-cadherin promoter in well and poorly differentiated carcinoma cell lines in order to gain insights into the mechanisms of E-cadherin down-regulation in tumors. In vivo footprinting analysis revealed that positive regulatory elements of the E-cadherin promoter (a GC-rich region, the CCAAT-box and a palindromic element) are specifically bound by transcription factors in E-cadherin-expressing but not in non-expressing cells. The tested cell systems include more than a dozen carcinomas cell lines as well as mammary epithelial cells where E-cadherin expression can be switched off by activation of a Fos-estrogen receptor fusion protein and rhabdomyosarcoma cells where E-cadherin expression was induced by transfection with E1A. Mapping of DNase I hypersensitive sites showed that the chromatin structure in the promoter region is loosened in expressing but condensed in non-expressing cells. Furthermore, the endogenous E-cadherin promoter is specifically methylated at CpG sites in the undifferentiated cells. We also show that the in vivo properties of the promoter in E-caherin-negative carcinoma cells are similar as in mesenchymal cells, i.e. fibroblasts or sarcoma cells. These data suggest that silencing of the E-cadherin promoter during epithelialmesenchymal transition and tumor progression is due to a loss of factor binding in vivo and to chromatin rearrangement in the regulatory region.

Animals↗
PubMed1995

Characterization of the scatter factor/hepatocyte growth factor gene promoter. Positive and negative regulatory elements direct gene expression to mesenchymal cells.

Scatter factor/hepatocyte growth factor (SF/HGF) and its receptor c-Met represent a paracrine signaling system involved in mesenchymal-epithelial interactions during development and during tumor progression. We have examined the promoters of the mouse and human SF/HGF genes by deletion mapping followed by CAT assays as well as by gel retardation and footprinting analysis. The promoter sequences are highly conserved (89.5% identity) up to position -453 from the major transcription start site but diverged considerably further upstream. Both promoters are active in mesenchymal but not epithelial cells thus reflecting the expression pattern of the SF/HGF gene in cells in vitro and in vivo. We have here identified two regulatory sequences in the SF/HGF promoter: a negative element at positions -239 to -258 and a positive element near the major transcription start site; specific deletions destroyed the activities of these elements. We were not able to localize elements on the SF/HGF promoter region that mediate the previously described effects of transforming growth factor beta, 12-O-tetradecanoylphorbol-13-acetate, and coculture of epithelial cells on SF/HGF gene expression. This study represents a first step toward understanding the intricately regulated and cell type-specific expression of the paracrine acting SF/HGF.

3T3 Cells↗
PubMed1995

E-cadherin as an invasion suppressor.

The loss of epithelial differentiation in carcinomas, which is accompanied by increased mobility and invasiveness of the tumour cells, is often a consequence of reduced intercellular adhesion. Recent reports have indicated that the primary cause for the 'scattering' of the cells in invasive carcinomas is a disturbance of the integrity of intercellular junctions often involving the cell adhesion molecule E-cadherin. It has also been suggested that during invasion, carcinoma cells convert to a sort of mesenchymal stage, as do normal epithelial cells during development. Permanent and transient molecular mechanisms lead to the impairment of junction integrity of epithelial cells and thus to the progression of carcinomas towards a more invasive state.

Cadherins↗
PubMed1995

Distribution of E-cadherin and Ep-CAM in the human lung during development and after injury.

Paraffin sections were obtained of human fetal, adult, and pathological lung (pulmonary fibrosis after radiotherapy or chemotherapy). The localization of epithelial adhesion molecules E-cadherin and Ep-CAM (former epithelial surface 40 kDa glycoprotein) was investigated by immunoperoxidase and/or immunofluorescence techniques with monoclonal antibodies. During development, the epithelia of the primary pulmonary primordium, the secondary bronchi and the adult bronchial epithelium retained immunoreactivity for E-cadherin and Ep-CAM with lateral immunostaining of cell membranes. In normal adult lungs, Ep-CAM was detected in type I and II alveolar epithelial cells, whereas E-cadherin was confined to the basolateral domain of type II cells. In pulmonary fibrosis, Ep-CAM could be further detected on the cell surface of epithelial remnants. In contrast, E-cadherin expression was characterized by a change of the membrane localization to a spotty, cytoplasmic pattern in the alveolar epithelium, possibly indicating functional inactivation of the protein during fibrogenesis.

Adult↗
PubMed1995

V-src kinase shifts the cadherin-based cell adhesion from the strong to the weak state and beta catenin is not required for the shift.

The elevation of tyrosine phosphorylation level is thought to induce the dysfunction of cadherin through the tyrosine phosphorylation of beta catenin. We evaluated this assumption using two cell lines. First, using temperature-sensitive v-src-transfected MDCK cells, we analyzed the modulation of cadherin-based cell adhesion by tyrosine phosphorylation. Cell aggregation and dissociation assays at nonpermissive and permissive temperatures indicated that elevation of the tyrosine phosphorylation does not totally affect the cell adhesion ability of cadherin but shifts it from a strong to a weak state. The tyrosine phosphorylation levels of beta catenin, ZO-1, ERM (ezrin/radixin/moesin), but not alpha catenin, vinculin, and alpha-actinin, were elevated in the weak state. To evaluate the involvement of the tyrosine phosphorylation of beta catenin in this shift of cadherin-based cell adhesion, we introduced v-src kinase into L fibroblasts expressing the cadherin-alpha catenin fusion protein, in which beta catenin is not involved in cell adhesion. The introduction of v-src kinase in these cells shifted their adhesion from a strong to a weak state. These findings indicated that the tyrosine phosphorylation of beta catenin is not required for the strong-to-weak state shift of cadherin-based cell adhesion, but that the tyrosine phosphorylation of other junctional proteins, ERM, ZO-1 or unidentified proteins is involved.

Animals↗
PubMed1995

Adherens junction proteins in tumour progression.

The loss of epithelial differentiation in carcinomas, which is accompanied by higher mobility and invasiveness of the tumour cells, is often a consequence of reduced intercellular adhesion. The primary cause of the "scattering" of cells in invasive carcinomas appears to be a disturbance of the integrity of intercellular junctions, often involving the cell adhesion molecule E-cadherin. Permanent and transient molecular mechanisms can lead to the impairment of junction integrity of epithelial cells and thus to the progression of carcinomas towards a more invasive state. These include downregulation of E-cadherin expression and interaction between the adherens junction protein beta-catenin and the tumour suppressor gene product APC.

Adenomatous Polyposis Coli Protein↗
PubMed1994

Localization of the human beta-catenin gene (CTNNB1) to 3p21: a region implicated in tumor development.

The human beta-catenin locus (CTNNB1) was mapped by in situ fluorescence analysis to band p21 on the short arm of chromosome 3, a region frequently affected by somatic alterations in a variety of tumors. PCR primers for the genomic amplification of beta-catenin sequences were selected on the basis of homology to exon 4 of the Drosophila armadillo gene. Analysis of a panel of somatic cell hybrids confirmed the localization of beta-catenin on human chromosome 3. Furthermore, exclusion mapping of three hybrids carrying defined fragments of the short arm of human chromosome 3 allowed us to determine the position of the CTNNB1 locus close to the marker D3S2 in 3p21.

Adenomatous Polyposis Coli Protein↗