title: RACK1 (receptor for activated C-kinase 1) interacts with FBW2 (F-box and WD-repeat domain-containing 2) to up-regulate GCM1 (glial cell missing 1) stability and placental cell migration and invasion.
abstract: GCM1 (glial cell missing 1) is a short-lived transcription factor essential for placental development. The F-box protein, FBW2 (F-box and WD-repeat domain-containing 2), which contains five WD (tryptophan-aspartate) repeats, recognizes GCM1 and mediates its ubiquitination via the SCFFBW2 E3 ligase complex. Although the interaction between GCM1 and FBW2 is facilitated by GCM1 phosphorylation, it is possible that this interaction might be regulated by additional cellular factors. In the present study, we perform tandem-affinity purification coupled with MS analysis identifying RACK1 (receptor for activated C-kinase 1) as an FBW2-interacting protein. RACK1 is a multifaceted scaffold protein containing seven WD repeats. We demonstrate that the WD repeats in both RACK1 and FBW2 are required for the interaction of RACK1 and FBW2. Furthermore, RACK1 competes with GCM1 for FBW2 and thereby prevents GCM1 ubiquitination, which is also supported by the observation that GCM1 is destabilized in RACK1-knockdown BeWo placental cells. Importantly, RACK1 knockdown leads to decreased expression of the GCM1 target gene HTRA4 (high-temperature requirement protein A4), which encodes a serine protease crucial for cell migration and invasion. As a result, migration and invasion activities are down-regulated in RACK1-knockdown BeWo cells. The present study reveals a novel function for RACK1 to regulate GCM1 activity and placental cell migration and invasion.
authors: Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei 115, Taiwan.
authors: Chang-Chun
authors: CC
authors: Wang
authors: Hsiao-Fan
authors: HF
authors: Lo
authors: Shu-Yu
authors: SY
authors: Lin
authors: Hungwen
authors: H
authors: Chen
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D002465:Cell Movement; D017931:DNA Primers; D004268:DNA-Binding Proteins; D044783:F-Box Proteins / Q000378:metabolism*; D005260:Female; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D010920:Placenta / Q000166:cytology / Q000378:metabolism*; D011247:Pregnancy; D060888:Real-Time Polymerase Chain Reaction; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D054875:Ubiquitination; D015854:Up-Regulation
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; D044783:F-Box Proteins; C499832:FBXW2 protein, human; C491089:GCM1 protein, human; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; C419368:RACK1 protein, human; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D014157:Transcription Factors; D019204:GTP-Binding Proteins