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TGF-beta induces the expression of SAP30L, a novel nuclear protein.

Lindfors K, Viiri KM, Niittynen M, Heinonen TY, Mäki M, Kainulainen H - BMC Genomics (2003)

Bottom Line: Differential display analysis resulted in the identification of a novel TGF-beta upregulated mRNA species, the Sin3-associated protein 30-like, SAP30L.The mRNA is expressed in several human tissues and codes for a nuclear protein of 183 amino acids 70% identical with Sin3 associated protein 30 (SAP30).The predicted nuclear localization signal of SAP30L is sufficient for nuclear transport of the protein although mutating it does not completely remove SAP30L from the nuclei.

View Article: PubMed Central - HTML - PubMed

Affiliation: Paediatric Research Centre, Tampere University Hospital, Tampere, Finland. katri.lindfors@uta.fi

ABSTRACT

Background: We have previously set up an in vitro mesenchymal-epithelial cell co-culture model which mimics the intestinal crypt villus axis biology in terms of epithelial cell differentiation. In this model the fibroblast-induced epithelial cell differentiation from secretory crypt cells to absorptive enterocytes is mediated via transforming growth factor-beta (TGF-beta), the major inhibitory regulator of epithelial cell proliferation known to induce differentiation in intestinal epithelial cells. The aim of this study was to identify novel genes whose products would play a role in this TGF-beta-induced differentiation.

Results: Differential display analysis resulted in the identification of a novel TGF-beta upregulated mRNA species, the Sin3-associated protein 30-like, SAP30L. The mRNA is expressed in several human tissues and codes for a nuclear protein of 183 amino acids 70% identical with Sin3 associated protein 30 (SAP30). The predicted nuclear localization signal of SAP30L is sufficient for nuclear transport of the protein although mutating it does not completely remove SAP30L from the nuclei. In the nuclei SAP30L concentrates in small bodies which were shown by immunohistochemistry to colocalize with PML bodies only partially.

Conclusions: By reason of its nuclear localization and close homology to SAP30 we believe that SAP30L might have a role in recruiting the Sin3-histone deacetylase complex to specific corepressor complexes in response to TGF-beta, leading to the silencing of proliferation-driving genes in the differentiating intestinal epithelial cells.

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The expression of SAP30L in control and TGF-β-treated differentiated T84 epithelial cells. The lower panel shows that in DD-PCR the band representing SAP30L was present solely in the RNA sample from the differentiated T84 cells. Real time quantitative PCR verified this differential expression to be 2.0 times higher (SEM ± 0.13).
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Figure 1: The expression of SAP30L in control and TGF-β-treated differentiated T84 epithelial cells. The lower panel shows that in DD-PCR the band representing SAP30L was present solely in the RNA sample from the differentiated T84 cells. Real time quantitative PCR verified this differential expression to be 2.0 times higher (SEM ± 0.13).

Mentions: DD-PCR showed that TGF-β1 induced consistent and reproducible upregulation of a transcript denoted SAP30L (see later) (Figure 1) using the arbitrary 5' primer AP-3 and the 3' anchoring primer T12MG. Quantitative RT-PCR using LightCycler technology verified this induction by TGF-β in three independent experiments. The differentiated TGF-β-treated cells expressed this transcript 2.0 times more than the unstimulated T84 cells (Figure 1).


TGF-beta induces the expression of SAP30L, a novel nuclear protein.

Lindfors K, Viiri KM, Niittynen M, Heinonen TY, Mäki M, Kainulainen H - BMC Genomics (2003)

The expression of SAP30L in control and TGF-β-treated differentiated T84 epithelial cells. The lower panel shows that in DD-PCR the band representing SAP30L was present solely in the RNA sample from the differentiated T84 cells. Real time quantitative PCR verified this differential expression to be 2.0 times higher (SEM ± 0.13).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC319701&req=5

Figure 1: The expression of SAP30L in control and TGF-β-treated differentiated T84 epithelial cells. The lower panel shows that in DD-PCR the band representing SAP30L was present solely in the RNA sample from the differentiated T84 cells. Real time quantitative PCR verified this differential expression to be 2.0 times higher (SEM ± 0.13).
Mentions: DD-PCR showed that TGF-β1 induced consistent and reproducible upregulation of a transcript denoted SAP30L (see later) (Figure 1) using the arbitrary 5' primer AP-3 and the 3' anchoring primer T12MG. Quantitative RT-PCR using LightCycler technology verified this induction by TGF-β in three independent experiments. The differentiated TGF-β-treated cells expressed this transcript 2.0 times more than the unstimulated T84 cells (Figure 1).

Bottom Line: Differential display analysis resulted in the identification of a novel TGF-beta upregulated mRNA species, the Sin3-associated protein 30-like, SAP30L.The mRNA is expressed in several human tissues and codes for a nuclear protein of 183 amino acids 70% identical with Sin3 associated protein 30 (SAP30).The predicted nuclear localization signal of SAP30L is sufficient for nuclear transport of the protein although mutating it does not completely remove SAP30L from the nuclei.

View Article: PubMed Central - HTML - PubMed

Affiliation: Paediatric Research Centre, Tampere University Hospital, Tampere, Finland. katri.lindfors@uta.fi

ABSTRACT

Background: We have previously set up an in vitro mesenchymal-epithelial cell co-culture model which mimics the intestinal crypt villus axis biology in terms of epithelial cell differentiation. In this model the fibroblast-induced epithelial cell differentiation from secretory crypt cells to absorptive enterocytes is mediated via transforming growth factor-beta (TGF-beta), the major inhibitory regulator of epithelial cell proliferation known to induce differentiation in intestinal epithelial cells. The aim of this study was to identify novel genes whose products would play a role in this TGF-beta-induced differentiation.

Results: Differential display analysis resulted in the identification of a novel TGF-beta upregulated mRNA species, the Sin3-associated protein 30-like, SAP30L. The mRNA is expressed in several human tissues and codes for a nuclear protein of 183 amino acids 70% identical with Sin3 associated protein 30 (SAP30). The predicted nuclear localization signal of SAP30L is sufficient for nuclear transport of the protein although mutating it does not completely remove SAP30L from the nuclei. In the nuclei SAP30L concentrates in small bodies which were shown by immunohistochemistry to colocalize with PML bodies only partially.

Conclusions: By reason of its nuclear localization and close homology to SAP30 we believe that SAP30L might have a role in recruiting the Sin3-histone deacetylase complex to specific corepressor complexes in response to TGF-beta, leading to the silencing of proliferation-driving genes in the differentiating intestinal epithelial cells.

Show MeSH
Related in: MedlinePlus