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J. Biol. Chem., Vol. 277, Issue 51, 50143-50154, December 20, 2002
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From the Renal-Electrolyte and Hypertension Division, Department of
Medicine, University of Pennsylvania School of Medicine,
Philadelphia, Pennsylvania 19104-6144
Alternative splicing of fibroblast growth factor
receptor 2 (FGFR2) mutually exclusive exons IIIb and IIIc represents a
tightly regulated and functionally relevant example of
post-transcriptional gene regulation. Rat prostate cancer DT3 and AT3
cell lines demonstrate exclusive selection of either exon IIIb or exon
IIIc, respectively, and have been used to characterize regulatory FGFR2
RNA cis-elements that are required for splicing regulation.
Two sequences termed ISE-2 and ISAR are located in the intron between
exons IIIb and IIIc and are required for cell-type specific exon IIIb.
Previous studies suggest that the function of these elements involves
formation of an RNA stem structure, even though they are separated by
more than 700 nucleotides. Using transfected minigenes, we performed a
systematic analysis of the sequence and structural components of ISE-2
and ISAR that are required for their ability to regulate FGFR2
splicing. We found that the primary sequence of these elements can be
replaced by completely unrelated sequences, provided that they are also
predicted to form an RNA stem structure. Thus, a nonsequence-specific
double stranded RNA stem constitutes a functional element
required for FGFR2 splicing; suggesting that a double-stranded RNA
binding protein is a component of the splicing regulatory machinery.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EBI Data Bank with accession number(s) AYI61008 and AYI61009.
A Non-sequence-specific Double-stranded RNA Structural Element
Regulates Splicing of Two Mutually Exclusive Exons of Fibroblast Growth
Factor Receptor 2 (FGFR2)*
*
This work was supported by start-up funds from the
University of Pennsylvania School of Medicine, Department of Defense
Grant PC991539, and United States Public Health Services Grant K08
CA72560 from the NCI, National Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: University of
Pennsylvania School of Medicine, 700 Clinical Research Bldg., 415 Curie
Blvd., Philadelphia, PA 19104-6144. Tel.: 215-573-1838; Fax:
215-898-0189; E-mail: russcars@mail.med.upenn.edu.
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