Construction of mutants of phosphorylation sites in ERα AF-1 and effects of mutants on transcriptional activity of ERα

Kang Ning, Ya Juan Liu, Huai Ning Yang, Hong Xue Fan, Zhi Hong Yang, Rong Li, Qi Nong Ye

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: To construct the mutants of the four serine phosphorylation sites (104, 106, 118 and 167) in ERα AF-1 and detect their effects on the transcriptional activity of Erα in 293T cells. Methods: The coding sequences of the ERα mutants were amplified by recombinant PCR with pcDNA3-ERα as a template and fused in frame with the coding region of FLAG in the pcDNA3-FLAG vector. The fusion proteins were characterized by Western blotting. The effects of the mutants on the transcriptional activity of ERα were determined. Results: ERα mutants were successfully constructed and expressed in 293T cells. The transcriptional activities of ERα and its mutants were 3.40, 3.21, 3.02, 3.00, 3.54 times of pcDNA3 in the absence of E2. Only the mutants of 118 and 167 phosphorylation sites made the transcriptional activity reduce to 50% of ERα in the presence of E2. Conclusion: Only 118 and 167 phosphorylation sites are necessary during the regulation of estrogen on target gene transcription among the four serine phosphorylation sites (104, 106, 118 and 167) in ERα AF1.

Original languageEnglish (US)
Pages (from-to)200-203
Number of pages4
JournalJournal of Jilin University Medicine Edition
Volume33
Issue number2
StatePublished - Mar 2007

Keywords

  • Estrogen
  • Point mutation
  • Receptors
  • Trans-activation (genetics)

ASJC Scopus subject areas

  • Pharmacology

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