Identification of FBXO25-interacting proteins using an integrated proteomics approach
journal contributionposted on 01.03.2011 by Felipe R. Teixeira, Sami Yokoo, Carlos G. Gartner, Adriana O. Manfiolli, Munira M. A. Baqui, Eliana M. Assmann, Ana Leticia G. C. Maragno, Yu Huijun, Primal de Lanerolle, Jörg Kobarg, Steven P. Gygi, Marcelo D. Gomes
Any type of content formally published in an academic journal, usually following a peer-review process.
FBXO25 is one of 68 human F-box proteins that serve as specificity factors for a family of ubiquitin ligases composed of Skp1, Rbx1, Cullin1 and F-box protein (SCF1) that are involved in targeting proteins for destruction across the ubiquitin proteasome system. We recently reported that the FBXO25 protein accumulates in novel subnuclear structures named FBXO25-associated nuclear domains (FANDs). Combining two-step affinity purification followed by mass spectrometry with a classical two-hybrid screen, we identified 132 novel potential FBXO25 interacting partners. One of the identified proteins, -actin, physically interacts through its N-terminus with FBXO25 and is enriched in the FBXO25 nuclear compartments. Inhibitors of actin polymerization promote a significant disruption of FANDs, indicating that they are compartments influenced by the organizational state of actin in the nucleus. Furthermore, FBXO25 antibodies interfered with RNA polymerase II transcription in vitro. Our results open new perspectives for the understanding of this novel compartment and its nuclear functions.