Transcriptional Activity of the Promoter Region of Rat Frizzled-Related Protein Gene

doi:10.1006/bbrc.2001.5349

Biochemical and Biophysical Research Communications

Volume 286, Issue 1, 10 August 2001, Pages 94-100

https://doi.org/10.1006/bbrc.2001.5349 Get rights and content

Abstract

Frizzled-related protein (Frp) is a new family of secreted proteins involved in tumorigenesis and Wnt-signaling pathway. Previous study has shown that rat Frp (rFrp) gene was found to be differentially expressed in Rat 6 fibroblast cell line overexpressing p53^val135 (R6#13-8). The rFrp gene was otherwise silent in normal parental Rat 6 cells. To elucidate the molecular basis of the transcriptional activation of rFrp, we have isolated and analyzed a 2-kilobase pair promoter region of the rFrp gene. Mapping of transcription initiation sites of rFrp showed the existence of multiple initiation sites. Transfection studies of serial deletion constructs in both Rat 6 and CHOK1 cell lines revealed that the region from −202 to −144 contains cis-acting elements essential for the efficient transcription of rFrp. This work provides a transcriptional regulation basis for Frp and gives insight into its implication in tumorigenesis.

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Cited by (7)

Genomic structure, alternative splicing and tissue expression of rFrp/sFRP-4, the rat frizzled related protein gene
2005, Gene
Secreted frizzled related proteins (sFRP) are regulators of Wnt signaling pathways that play central roles in developmental processes and oncogenesis. Various sFRP genes have been cloned from different tissues and implicated in diverse biological activities. rFrp, the rat homologue of sFRP-4, was initially identified as being upregulated in mutant p53-induced cellular transformation. Here, we report on the isolation of five novel splice variants, rFrp/sFRP-4 II, II, III, IVa and IVb. The complete rFrp/sFRP-4 genomic structure spans over 31 kb covering 9 exons. Except for the variant IVb, which was derived from IVa by alternative polyadenylation signal, variants I to IVa were alternatively spliced to different exons in the 3'end of mRNA and resulted in transcripts with truncated open reading frame. The deduced proteins of the variants had truncated C-termini, however, the two key functional protein domains, the cysteine-rich domain and the netrin-like domain of the isoforms, were not altered. In addition, different transcriptional initiation sites were found with variants II and IV, implying that these variants may be regulated differently from the rFrp/sFRP-4. RT-PCR analysis showed that these splice variants displayed different patterns of tissue-specific expression. Northern blot analysis revealed that the rFrp/sFRP-4 is most abundant in the ovary. Taken together, our findings suggest that alternative splicing of rFrp/sFRP-4 plays a role in regulating tissue-specific expression. The truncated C terminals of rFrp/sFRP-4 variants may confer structural specificity and hence exert different biological functions in different tissues. Characterization of these novel splice variants should help to elucidate the function of the sFRP family gene.
Increased expression of secreted frizzled-related protein 4 in polycystic kidneys
2009, Journal of the American Society of Nephrology
SFRP-4 abrogates Wnt-3a-induced β-catenin and Akt/PKB signalling and reverses a Wnt-3a-imposed inhibition of in vitro mammary differentiation
2008, Journal of Molecular Signaling
Expression and localization of secreted frizzled-related protein-4 in the rodent ovary: Evidence for selective up-regulation in luteinized granulosa cells
2003, Endocrinology
Cloning and characterization of the promoter region of the mouse frizzled-related protein 4 gene
2003, Biological Chemistry
Transcriptional regulation of the promoter of the rat frizzled related protein gene by CREB
2003, Oncogene

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^☆: Sequence information is deposited with the GenBank Accession No. AF140346, AF140347, and AF364906.

^☆☆: Abbreviations used: rFrp, rat frizzled-related protein; Fz, frizzled; CDR, cysteine-rich domain; RACE, rapid amplification of cDNA ends; SRY, sex-determining region Y gene product; STAT, signal transducer and activator of transcription; MZF, myeloid zinc finger; C/EBP-β, CCAAT/enhancer binding protein beta; CREB, cAMP responsive element binding protein; CBP, CREB binding protein; IK2, Ikaros 2.

¹: To whom correspondence should be addressed. Fax: (852) 2358-1559. E-mail: [email protected].

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Biochemical and Biophysical Research Communications

Abstract

References (27)

Primary structure and tissue distribution of Frzb, a novel protein related to Drosophila frizzled, suggest a role in skeletal morphogenesis

J. Biol. Chem.

Frzb-1 is a secreted antagonist of Wnt signaling expressed in the spemann organizer

Cell

Frzb, a secreted protein expressed in the spemann organizer, binds and inhibits Wnt-8

Cell

Interaction of frizzled related protein (FRP) with wnt ligands and the frizzled receptor suggests alternative Mechanisms for FRP Inhibition of Wnt Signaling

J. Biol. Chem.

Secreted frizzled-related protein-1 binds directly to wingless and is a biphasic modulator of wnt signaling

J. Biol. Chem.

DDC-4, an apoptosis-associated gene, is a secreted frizzled relative

FEBS Lett.

Identification and characterization of genes whose expressions are altered in rat 6 fibroblasts transformed by mutant p53^val135

Biochem. Biophy. Res. Commun.

Solution structure of the KIX domain of CBP bound to the transactivation domain of CREB: A model for activator:coactivator interactions

Cell

Binding of p53 to the KIX domain of CREB binding protein. A potential link to human T-cell leukamia virus, type I-associated leukemogenesis

J. Biol. Chem.

A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors

Proc. Natl. Acad. Sci. USA

Purification and molecular cloning of a secreted, frizzled-related antagonist of Wnt action

Proc. Natl. Acad. Sci. USA

A new member of the frizzled family from Drosophila functions as a Wingless receptor

Nature

Activation of β-catenin-Tcf signaling in colon cancer by mutations in β-catenin of APC

Science

Cited by (7)

Genomic structure, alternative splicing and tissue expression of rFrp/sFRP-4, the rat frizzled related protein gene

Increased expression of secreted frizzled-related protein 4 in polycystic kidneys

SFRP-4 abrogates Wnt-3a-induced β-catenin and Akt/PKB signalling and reverses a Wnt-3a-imposed inhibition of in vitro mammary differentiation

Expression and localization of secreted frizzled-related protein-4 in the rodent ovary: Evidence for selective up-regulation in luteinized granulosa cells

Cloning and characterization of the promoter region of the mouse frizzled-related protein 4 gene

Transcriptional regulation of the promoter of the rat frizzled related protein gene by CREB

Regular ArticleTranscriptional Activity of the Promoter Region of Rat Frizzled-Related Protein Gene☆,☆☆

Abstract

J. Biol. Chem.

Cell

Cell

J. Biol. Chem.

J. Biol. Chem.

FEBS Lett.

Biochem. Biophy. Res. Commun.

Cell

J. Biol. Chem.

A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors

Proc. Natl. Acad. Sci. USA

Purification and molecular cloning of a secreted, frizzled-related antagonist of Wnt action

Proc. Natl. Acad. Sci. USA

A new member of the frizzled family from Drosophila functions as a Wingless receptor

Nature

Activation of β-catenin-Tcf signaling in colon cancer by mutations in β-catenin of APC

Science

Regular Article
Transcriptional Activity of the Promoter Region of Rat Frizzled-Related Protein Gene☆,☆☆