Identification and characterization of novel proteins associated with CHD4.

Genes Cells

Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.

Published: January 2022

AI Article Synopsis

  • The CHD family contains nine ATP-dependent chromatin remodeling factors, with CHD4 playing a crucial role in regulating various cellular activities through interactions with several proteins, including the NuRD complex.
  • New research identified LCORL and NOL4L as novel proteins interacting with CHD4, using FLAG-tagged CHD4 in HeLa-S3 and HEK293T cells.
  • RNA-sequencing data indicated that depleting CHD4, LCORL, or NOL4L led to the up-regulation of genes associated with the Notch signaling pathway, suggesting these proteins collaborate with CHD4 to suppress this pathway in mammalian cells.

Article Abstract

The CHD (chromodomain helicase DNA binding protein) family consists of nine chromatin remodeling factors that alter chromatin structure in an ATP-dependent manner. CHD4 contributes to the regulation of various cellular activities and processes including development through interaction with multiple proteins including formation of the NuRD (nucleosome remodeling and deacetylase activity) complex. Functions of CHD4 that appear not to be mediated by the NuRD complex or other known interactors have also been identified, however, suggesting the existence of unrecognized proteins that also associate with CHD4. We here generated HeLa-S3 and HEK293T cells with a knock-in allele for FLAG epitope-tagged CHD4 and used these cells to identify proteins that bind to CHD4 with the use of immunoprecipitation followed by liquid chromatography and tandem mass spectrometry. LCORL (ligand-dependent nuclear receptor corepressor like) and NOL4L (nucleolar protein 4 like) were reproducibly identified as novel CHD4 interactors. Furthermore, RNA-sequencing analysis of HEK293T cells depleted of CHD4, LCORL, or NOL4L revealed consistent up-regulation of genes related to the Notch signaling pathway. Our results thus suggest that both LCORL and NOL4L may cooperate with CHD4 to suppress the Notch pathway in mammalian cells.

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Source
http://dx.doi.org/10.1111/gtc.12909DOI Listing

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