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Alterations in cell growth and signaling in ErbB3 binding protein-1 (Ebp1) deficient mice.

Zhang Y, Lu Y, Zhou H, Lee M, Liu Z, Hassel BA, Hamburger AW - BMC Cell Biol. (2008)

Bottom Line: The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells.Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway.These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USA. yzhan001@umaryland.edu

ABSTRACT

Background: The ErbB3 binding protein-1 (Ebp1) belongs to a family of DNA/RNA binding proteins implicated in cell growth, apoptosis and differentiation. However, the physiological role of Ebp1 in the whole organism is not known. Therefore, we generated Ebp1-deficient mice carrying a gene trap insertion in intron 2 of the Ebp1 (pa2g4) gene.

Results: Ebp1-/- mice were on average 30% smaller than wild type and heterozygous sex matched littermates. Growth retardation was apparent from Day 10 until Day 30. IGF-1 production and IGBP-3 and 4 protein levels were reduced in both embryo fibroblasts and adult knock-out mice. The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells. Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway. In addition, the expression or activation of proliferation related genes such as AKT and the androgen receptor, previously demonstrated to be affected by Ebp1 expression in vitro, was altered in adult tissues.

Conclusion: These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific. The Ebp1-/- mouse line represents a new in vivo model to investigate Ebp1 function in the whole organism.

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Related in: MedlinePlus

Generation of the Ebp1  mouse. A. Schematic representation of the retroviral integration site within the first intron of the mouse pa2g4 (Ebp1) gene in the ES clone OST 186047 is indicated. Exons are represented as boxes. B. The locations of the PCR primers (LTR2, Ebp1 R, Ebp1 F) used in genotyping are indicated. Their positions (in base pairs) relative to the sites of integration are shown (not drawn to scale). LTR = long terminal repeat; SA and SD = splice acceptor and splice donor sites, respectively; PGK = phosphoglycerate kinase 1. C. A typical genotyping analysis showing the 308 bp PCR fragment for a wild type allele and the 253 bp product that detects the viral integration site. The products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.
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Figure 1: Generation of the Ebp1 mouse. A. Schematic representation of the retroviral integration site within the first intron of the mouse pa2g4 (Ebp1) gene in the ES clone OST 186047 is indicated. Exons are represented as boxes. B. The locations of the PCR primers (LTR2, Ebp1 R, Ebp1 F) used in genotyping are indicated. Their positions (in base pairs) relative to the sites of integration are shown (not drawn to scale). LTR = long terminal repeat; SA and SD = splice acceptor and splice donor sites, respectively; PGK = phosphoglycerate kinase 1. C. A typical genotyping analysis showing the 308 bp PCR fragment for a wild type allele and the 253 bp product that detects the viral integration site. The products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.

Mentions: Ebp1+/- mice were generated at Lexicon Genetics from ES cells containing a retroviral gene trap in the Ebp1 locus as described previously [23]. Sequence analysis showed that the gene trap was inserted at a single site in the 2.3 kb intron 2 of the Ebp1 gene (Fig. 1A, B). The insertion led to the expression of an Ebp1-neomycin fusion molecule containing only the first 20 amino acids that are encoded by Exon 1 of Ebp1 that has no known biological function. The mice were genotyped by PCR with primers located at each side of the gene trap and in the LTR2 cassette (Fig. 1C).


Alterations in cell growth and signaling in ErbB3 binding protein-1 (Ebp1) deficient mice.

Zhang Y, Lu Y, Zhou H, Lee M, Liu Z, Hassel BA, Hamburger AW - BMC Cell Biol. (2008)

Generation of the Ebp1  mouse. A. Schematic representation of the retroviral integration site within the first intron of the mouse pa2g4 (Ebp1) gene in the ES clone OST 186047 is indicated. Exons are represented as boxes. B. The locations of the PCR primers (LTR2, Ebp1 R, Ebp1 F) used in genotyping are indicated. Their positions (in base pairs) relative to the sites of integration are shown (not drawn to scale). LTR = long terminal repeat; SA and SD = splice acceptor and splice donor sites, respectively; PGK = phosphoglycerate kinase 1. C. A typical genotyping analysis showing the 308 bp PCR fragment for a wild type allele and the 253 bp product that detects the viral integration site. The products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2648959&req=5

Figure 1: Generation of the Ebp1 mouse. A. Schematic representation of the retroviral integration site within the first intron of the mouse pa2g4 (Ebp1) gene in the ES clone OST 186047 is indicated. Exons are represented as boxes. B. The locations of the PCR primers (LTR2, Ebp1 R, Ebp1 F) used in genotyping are indicated. Their positions (in base pairs) relative to the sites of integration are shown (not drawn to scale). LTR = long terminal repeat; SA and SD = splice acceptor and splice donor sites, respectively; PGK = phosphoglycerate kinase 1. C. A typical genotyping analysis showing the 308 bp PCR fragment for a wild type allele and the 253 bp product that detects the viral integration site. The products were separated by agarose gel electrophoresis and visualized by ethidium bromide staining.
Mentions: Ebp1+/- mice were generated at Lexicon Genetics from ES cells containing a retroviral gene trap in the Ebp1 locus as described previously [23]. Sequence analysis showed that the gene trap was inserted at a single site in the 2.3 kb intron 2 of the Ebp1 gene (Fig. 1A, B). The insertion led to the expression of an Ebp1-neomycin fusion molecule containing only the first 20 amino acids that are encoded by Exon 1 of Ebp1 that has no known biological function. The mice were genotyped by PCR with primers located at each side of the gene trap and in the LTR2 cassette (Fig. 1C).

Bottom Line: The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells.Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway.These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, University of Maryland School of Medicine, Baltimore, MD, USA. yzhan001@umaryland.edu

ABSTRACT

Background: The ErbB3 binding protein-1 (Ebp1) belongs to a family of DNA/RNA binding proteins implicated in cell growth, apoptosis and differentiation. However, the physiological role of Ebp1 in the whole organism is not known. Therefore, we generated Ebp1-deficient mice carrying a gene trap insertion in intron 2 of the Ebp1 (pa2g4) gene.

Results: Ebp1-/- mice were on average 30% smaller than wild type and heterozygous sex matched littermates. Growth retardation was apparent from Day 10 until Day 30. IGF-1 production and IGBP-3 and 4 protein levels were reduced in both embryo fibroblasts and adult knock-out mice. The proliferation of fibroblasts derived from Day 12.5 knock out embryos was also decreased as compared to that of wild type cells. Microarray expression analysis revealed changes in genes important in cell growth including members of the MAPK signal transduction pathway. In addition, the expression or activation of proliferation related genes such as AKT and the androgen receptor, previously demonstrated to be affected by Ebp1 expression in vitro, was altered in adult tissues.

Conclusion: These results indicate that Ebp1 can affect growth in an animal model, but that the expression of proliferation related genes is cell and context specific. The Ebp1-/- mouse line represents a new in vivo model to investigate Ebp1 function in the whole organism.

Show MeSH
Related in: MedlinePlus