To determine the characteristics of the N-terminal transactivation domain (AF-1) of the mouse estrogen receptor (ER), we constructed a number of deletion mutants. Wild-type and mutant receptors were expressed in yeast cells and assayed for their ability to transactivate an estrogen-responsive reporter plasmid (ERE-CYCl-LacZ) that contained a single estrogen response element of the vitellogenin A2 gene promoter. Deletion of the N-terminal 121 animo acids from the mouse ER resulted in a 50% reduction in transaction activity compared with the full-length wild-type ER. Deletion of the first 150 amino acids resulted in loss of 90% transactivation activity. An ER deletion mutant lacking residues 121-154 retained full transcriptional activity, suggesting that this region plays a significant transacting role only when the first portion is deleted. A point mutation was introduced in the C-terminal region at Met-521 in order to study the possible interaction between the C-terminal ligand-binding domain and the N-terminal AF-1 region. This mutant ER, M521G, exhibited 150% of the transcriptional activity of the wild-type ER. An M521G mutant lacking the N-terminal 121 amino acids retained full transactivation activity, whereas, M521G lacking 150 amino acids resulted in only 10% of wild-type activity. These results suggest that residues 121-154 might interact with the C terminus to affect transcription. In summary, multiple N-terminal regions in the ER were identified that function in transactivation. Furthermore, a point mutation in the C-terminal portion of the ER may change the conformation of the ER ligand-binding domain, producing a more stable receptor/ligand complex that increases transcriptional activity. These data suggest that the N- and C-terminal portions of the ER interact in a cooperative manner to activate transcription from target genes. Published by Elsevier Science Inc.

Two transcription activation functions in the amino terminus of the mouse estrogen receptor that are affected by the carboxy terminus / Gandini, Orietta; Hirao, Kohno; Sylvia, Curtis; Kenneth S., Korach. - In: STEROIDS. - ISSN 0039-128X. - 62:7(1997), pp. 508-515. [10.1016/s0039-128x(97)00001-9]

Two transcription activation functions in the amino terminus of the mouse estrogen receptor that are affected by the carboxy terminus

GANDINI, Orietta;
1997

Abstract

To determine the characteristics of the N-terminal transactivation domain (AF-1) of the mouse estrogen receptor (ER), we constructed a number of deletion mutants. Wild-type and mutant receptors were expressed in yeast cells and assayed for their ability to transactivate an estrogen-responsive reporter plasmid (ERE-CYCl-LacZ) that contained a single estrogen response element of the vitellogenin A2 gene promoter. Deletion of the N-terminal 121 animo acids from the mouse ER resulted in a 50% reduction in transaction activity compared with the full-length wild-type ER. Deletion of the first 150 amino acids resulted in loss of 90% transactivation activity. An ER deletion mutant lacking residues 121-154 retained full transcriptional activity, suggesting that this region plays a significant transacting role only when the first portion is deleted. A point mutation was introduced in the C-terminal region at Met-521 in order to study the possible interaction between the C-terminal ligand-binding domain and the N-terminal AF-1 region. This mutant ER, M521G, exhibited 150% of the transcriptional activity of the wild-type ER. An M521G mutant lacking the N-terminal 121 amino acids retained full transactivation activity, whereas, M521G lacking 150 amino acids resulted in only 10% of wild-type activity. These results suggest that residues 121-154 might interact with the C terminus to affect transcription. In summary, multiple N-terminal regions in the ER were identified that function in transactivation. Furthermore, a point mutation in the C-terminal portion of the ER may change the conformation of the ER ligand-binding domain, producing a more stable receptor/ligand complex that increases transcriptional activity. These data suggest that the N- and C-terminal portions of the ER interact in a cooperative manner to activate transcription from target genes. Published by Elsevier Science Inc.
1997
gene regulation; hormone; mutagenesis; receptor mutant; transcription
01 Pubblicazione su rivista::01a Articolo in rivista
Two transcription activation functions in the amino terminus of the mouse estrogen receptor that are affected by the carboxy terminus / Gandini, Orietta; Hirao, Kohno; Sylvia, Curtis; Kenneth S., Korach. - In: STEROIDS. - ISSN 0039-128X. - 62:7(1997), pp. 508-515. [10.1016/s0039-128x(97)00001-9]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/412617
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