Abstract
An insulin-like growth factor (IGF) I response element (IGFRE) in the porcine P-450 cholesterol side-chain cleavage gene (P450scc) regulates transcription through the binding of two proteins, Sp1 and polypyrimidine tract-binding protein-associated splicing factor (PSF). PSF is a component of spliceosomes and contains RNA-binding domains. In this study, we localized the NH2-terminal amino acid residues necessary for binding of PSF to the IGFRE. Three COOH-terminal truncated proteins (aa 304, 214, and 134) of PSF were designed to empirically partition the NH2-terminal region while excluding the RNA-binding domains. Southwestern analysis showed that only the largest expressed truncated protein, P3, strongly bound the porcine P450scc IGFRE. Truncated PSF protein expression in Y1 adrenal cells showed that P3 repressed transcriptional activity of the IGFRE similar to full-length PSF, whereas P2 (minimal binding to the IGFRE) had no effect. In conclusion, the NH2-terminal region of PSF contains the amino acid residues necessary for binding to the porcine P450scc IGFRE and repressing the transcriptional activity of the element.
| Original language | English (US) |
|---|---|
| Journal | American Journal of Physiology - Endocrinology and Metabolism |
| Volume | 283 |
| Issue number | 3 46-3 |
| State | Published - Sep 1 2002 |
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Keywords
- Insulin-like growth factor-I
- P-450 cholesterol side-chain cleavage
- Polypyrimidine tract-binding protein
- Sp1
- Steroidogenesis
- Transcription
ASJC Scopus subject areas
- Physiology
- Endocrinology
- Biochemistry
Cite this
NH2 terminus of PTB-associated splicing factor binds to the porcine P450scc IGF-I response element. / Urban, Randall; Bodenburg, Yvonne H.; Wood, Thomas.
In: American Journal of Physiology - Endocrinology and Metabolism, Vol. 283, No. 3 46-3, 01.09.2002.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - NH2 terminus of PTB-associated splicing factor binds to the porcine P450scc IGF-I response element
AU - Urban, Randall
AU - Bodenburg, Yvonne H.
AU - Wood, Thomas
PY - 2002/9/1
Y1 - 2002/9/1
N2 - An insulin-like growth factor (IGF) I response element (IGFRE) in the porcine P-450 cholesterol side-chain cleavage gene (P450scc) regulates transcription through the binding of two proteins, Sp1 and polypyrimidine tract-binding protein-associated splicing factor (PSF). PSF is a component of spliceosomes and contains RNA-binding domains. In this study, we localized the NH2-terminal amino acid residues necessary for binding of PSF to the IGFRE. Three COOH-terminal truncated proteins (aa 304, 214, and 134) of PSF were designed to empirically partition the NH2-terminal region while excluding the RNA-binding domains. Southwestern analysis showed that only the largest expressed truncated protein, P3, strongly bound the porcine P450scc IGFRE. Truncated PSF protein expression in Y1 adrenal cells showed that P3 repressed transcriptional activity of the IGFRE similar to full-length PSF, whereas P2 (minimal binding to the IGFRE) had no effect. In conclusion, the NH2-terminal region of PSF contains the amino acid residues necessary for binding to the porcine P450scc IGFRE and repressing the transcriptional activity of the element.
AB - An insulin-like growth factor (IGF) I response element (IGFRE) in the porcine P-450 cholesterol side-chain cleavage gene (P450scc) regulates transcription through the binding of two proteins, Sp1 and polypyrimidine tract-binding protein-associated splicing factor (PSF). PSF is a component of spliceosomes and contains RNA-binding domains. In this study, we localized the NH2-terminal amino acid residues necessary for binding of PSF to the IGFRE. Three COOH-terminal truncated proteins (aa 304, 214, and 134) of PSF were designed to empirically partition the NH2-terminal region while excluding the RNA-binding domains. Southwestern analysis showed that only the largest expressed truncated protein, P3, strongly bound the porcine P450scc IGFRE. Truncated PSF protein expression in Y1 adrenal cells showed that P3 repressed transcriptional activity of the IGFRE similar to full-length PSF, whereas P2 (minimal binding to the IGFRE) had no effect. In conclusion, the NH2-terminal region of PSF contains the amino acid residues necessary for binding to the porcine P450scc IGFRE and repressing the transcriptional activity of the element.
KW - Insulin-like growth factor-I
KW - P-450 cholesterol side-chain cleavage
KW - Polypyrimidine tract-binding protein
KW - Sp1
KW - Steroidogenesis
KW - Transcription
UR - http://www.scopus.com/inward/record.url?scp=0036710172&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036710172&partnerID=8YFLogxK
M3 - Article
C2 - 12169434
AN - SCOPUS:0036710172
VL - 283
JO - American Journal of Physiology - Endocrinology and Metabolism
JF - American Journal of Physiology - Endocrinology and Metabolism
SN - 0193-1849
IS - 3 46-3
ER -