title: Identification of FBXO25-interacting proteins using an integrated proteomics approach.
abstract: FBXO25 is one of the 68 human F-box proteins that serve as specificity factors for a family of ubiquitin ligases composed of s-phase-kinase associated protein 1, really interesting new gene-box 1, Cullin 1, 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 (FAND). Combining two-step affinity purification followed by MS with a classical two-hybrid screen, we identified 132 novel potential FBXO25 interacting partners. One of the identified proteins, beta-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 FAND, 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.
authors.affiliation: Department of Biochemistry and Immunology, University of São Paulo, Ribeirão Preto, Brazil.
authors.forename: Felipe R
authors.initials: FR
authors.lastname: Teixeira
authors.forename: Sami
authors.initials: S
authors.lastname: Yokoo
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Gartner
authors.forename: Adriana O
authors.initials: AO
authors.lastname: Manfiolli
authors.forename: Munira M A
authors.initials: MM
authors.lastname: Baqui
authors.forename: Eliana M
authors.initials: EM
authors.lastname: Assmann
authors.forename: Ana Leticia G C
authors.initials: AL
authors.lastname: Maragno
authors.forename: Huijun
authors.initials: H
authors.lastname: Yu
authors.forename: Primal
authors.initials: P
authors.lastname: de Lanerolle
authors.forename: Jörg
authors.initials: J
authors.lastname: Kobarg
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.forename: Marcelo Damário
authors.initials: MD
authors.lastname: Gomes
mesh_terms: D000199:Actins / Q000032:analysis / Q000378:metabolism; D000595:Amino Acid Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D044783:F-Box Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
chemical_list: D000199:Actins; D044783:F-Box Proteins; C510759:FBXO25 protein, human; D009419:Nerve Tissue Proteins; D020543:Proteome
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