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Protein Interactions and Structure Studies of the Transcriptional Modulator Ashwin
Author(s) -
Uzman Akif,
Lou ChihHong,
Fryszczyn Bartlomiej,
Pereida Norma,
Sater Amy
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.660.10
Subject(s) - chromatin immunoprecipitation , biology , xenopus , computational biology , lac repressor , gene , microbiology and biotechnology , promoter , gene expression , genetics , repressor
Ashwin is a novel modulator of canonical wnt‐dependent transcription, which regulates Organizer gene expression in Xenopus embryos. Coimmunoprecipitation studies demonstrate that Ashwin interacts with TCF3 in vivo, and that increasing amounts of Ashwin disrupt the interaction between beta‐catenin and TCF3. Chromatin immunoprecipitation studies indicate that overexpression of Ashwin reduces occupancy of the siamois promoter by beta‐catenin. Ashwin also interacts with Polycomb1 (Pc1), a member of the Polycomb Repressor Complex1 (PRC1), and our results suggest that Pc1 acts with Ashwin to inhibit beta‐catenin‐dependent gene expression. To learn more about the structure of Ashwin, we have used the Studier method of auto‐induction of recombinant protein under the control of lacI to obtain recombinant protein for crystallization and structure determination via X‐ray diffraction. Further, we present bioinformatic evidence that ashwin is a novel and highly conserved vertebrate gene that may have evolved from previously undescribed open reading frames