Identification and functional characterization of a bipartite nuclear localization signal in ANKRD11

Biochem Biophys Res Commun. 2023 Aug 30:670:117-123. doi: 10.1016/j.bbrc.2023.05.046. Epub 2023 May 15.

Abstract

ANKRD11 gene encodes for the large nuclear protein essential for multiple system development including the nervous system. However, the molecular basis for the proper nuclear localization of ANKRD11 has not yet been elucidated. In this study, we have identified a functional bipartite nuclear localization signal (bNLS) between residues 53 and 87 of ANKRD11. Using biochemical approaches, we discovered two major binding sites in this bipartite NLS for Importin α1. Through site-directed mutagenesis and functional analysis, we further found that this bipartite NLS is sufficient for nuclear import of overexpressing GFP in HeLa cells and necessary for nuclear localization of ANKRD11. Importantly, our study provides a possible pathogenic mechanism for certain clinical variants located within the bipartite nuclear localization signal of ANKRD11.

Keywords: ANKRD11; Bipartite nuclear localization signal (bNLS); Nuclear import.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Active Transport, Cell Nucleus
  • Amino Acid Sequence
  • Cell Nucleus / metabolism
  • HeLa Cells
  • Humans
  • Nuclear Localization Signals* / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Transcription Factors / metabolism
  • alpha Karyopherins* / metabolism

Substances

  • Nuclear Localization Signals
  • alpha Karyopherins
  • Transcription Factors
  • ANKRD11 protein, human
  • Repressor Proteins