Squamous cell carcinoma-related oncogene (SCCRO) neddylates Cul3 protein to selectively promote midbody localization and activity of Cul3KLHL21 protein complex during abscission

J Biol Chem. 2017 Sep 15;292(37):15254-15265. doi: 10.1074/jbc.M117.778530. Epub 2017 Jun 15.

Abstract

Squamous cell carcinoma-related oncogene (SCCRO)/DCUN1D1, a component of the neddylation E3 complex, regulates the activity of the cullin-RING-ligase type of ubiquitination E3s by promoting neddylation of cullin family members. Studies have shown that SCCRO regulates proliferation in vitro and in vivo Here we show that inactivation of SCCRO results in prolonged mitotic time because of delayed and/or failed abscission. The effects of SCCRO on abscission involve its role in neddylation and localization of Cul3 to the midbody. The Cul3 adaptor KLHL21 mediates the effects of SCCRO on abscission, as it fails to localize to the midbody in SCCRO-deficient cells during abscission, and its inactivation resulted in phenotypic changes identical to SCCRO inactivation. Ubiquitination-promoted turnover of Aurora B at the midbody was deficient in SCCRO- and KLHL21-deficient cells, suggesting that it is the target of Cul3KLHL21 at the midbody. Correction of abscission delays in SCCRO-deficient cells with addition of an Aurora B inhibitor at the midbody stage suggests that Aurora B is the target of SCCRO-promoted Cul3KLHL21 activity. The activity of other Cul3-anchored complexes, including Cul3KLHL9/KLHL13, was intact in SCCRO-deficient cells, suggesting that SCCRO selectively, rather than collectively, neddylates cullins in vivo Combined, these findings support a model in which the SCCRO, substrate, and substrate adaptors cooperatively provide tight control of neddylation and cullin-RING-ligase activity in vivo.

Keywords: cell cycle; inhibitor; mitosis; substrate specificity; ubiquitylation (ubiquitination).

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Aurora Kinase B / genetics
  • Aurora Kinase B / metabolism
  • Biomarkers / metabolism
  • Cell Cycle Proteins
  • Cells, Cultured
  • Cullin Proteins / chemistry
  • Cullin Proteins / genetics
  • Cullin Proteins / metabolism*
  • Cytoskeletal Proteins
  • Embryo, Mammalian / cytology
  • Intracellular Signaling Peptides and Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Knockout
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Microscopy, Confocal
  • Mutation
  • NEDD8 Protein
  • Protein Multimerization
  • Protein Processing, Post-Translational*
  • Protein Transport
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism*
  • RNA Interference
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Telophase
  • Time-Lapse Imaging
  • Ubiquitins / metabolism*

Substances

  • Biomarkers
  • Cell Cycle Proteins
  • Cul3 protein, mouse
  • Cullin Proteins
  • Cytoskeletal Proteins
  • DCUN1D1 protein, mouse
  • Intracellular Signaling Peptides and Proteins
  • Klhl21 protein, mouse
  • Luminescent Proteins
  • Microfilament Proteins
  • NEDD8 Protein
  • Nedd8 protein, mouse
  • Proto-Oncogene Proteins
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Ubiquitins
  • Aurkb protein, mouse
  • Aurora Kinase B