ITSN1 binds the E2-conjugating enzyme UBC9

  • K. O. Kozyrieva Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03143, Kyiv, Akad. Zabolotnogo str., 150
  • T. A. Gryaznova Institute of Molecular Biology and Genetics, NAS of Ukraine, Ukraine, 03143, Kyiv, Zabolotnogo str., 150 https://orcid.org/0000-0002-6479-4861
DOI: https://doi.org/10.7124/FEEO.v34.1612
Keywords: ITSN1 (intersectin 1), UBC9, SUMOylation, protein-protein interactions

Abstract

Aim. Scaffolding protein of the intersectin 1 (ITSN1) associated with malignant cell transformation. A short isoform of ITSN1 (ITSN1-S) can localize to the nucleus and inhibit breast cancer cell proliferation but the exact mechanisms of ITSN1 nuclear export have not been fully elucidated. SUMOylation of ITSN1, or its interaction with components of SUMO modification, may be one of the regulatory mechanisms contributing to the nuclear-cytoplasmic shuffle of ITSN1 in the cell. Methods. Full-length human UBC9 sequence was subcloned in pGEX4T2 vector for in vitro GST-binding assays with overexpressed Omni-ITSN1-S in 293 cell line. Lysates of 293 cells with overexpressed FLAG-UBC9 were used for co-immunoprecipitation with endogenous proteins of ITSN1 and ITSN2. Results. Endogenous ITSN1-S form complexes with full-length overexpressed UBC9 in 293 in vivo. Further analysis revealed that GST-UBC9 binds human full-length short isoform ITSN1-S in vitro. Conclusions. E2-conjugating enzyme of the SUMOylation, UBC9, is confirmed as a novel protein partner for ITSN1 both in vitro and in vivo. Considering the tumor suppressor role of a nuclear ITSN1-S in breast cancer and the unique role UBC9 plays in SUMO-modification of proteins, we suggest a possibility of UBC9 and ITSN1 interaction association with malignant transformation, which can be the ground for the further studies.

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