Identification of circNLK-interacting proteins by RNA pull-down coupled with LC-MS/MS in glioblastoma cells

ABSTRACT: Glioblastoma is an aggressive primary brain malignancy with a poor prognosis. In this study, we investigated the molecular function of circNLK (hsa_circ_0002103), a circular RNA that is downregulated in glioblastoma and associated with improved patient survival. To identify proteins interacting with circNLK, RNA pull-down assays were performed using a biotin-labeled circNLK-specific probe and a corresponding control probe, followed by liquid chromatography-tandem mass spectrometry analysis. Mass spectrometry identified hnRNP L as a candidate circNLK-interacting RNA-binding protein. Subsequent RNA pull-down and RNA immunoprecipitation assays confirmed the interaction between circNLK and hnRNP L. Functional studies supported a model in which circNLK competitively binds hnRNP L in an m6A-sensitive manner, thereby reducing hnRNP L binding to the 3′ untranslated region of RGS16 mRNA, decreasing RGS16 mRNA stability and expression, and suppressing glioblastoma progression.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Brain, Permanent Cell Line Cell

DISEASE(S): Brain Cancer

SUBMITTER: xue kong  

LAB HEAD: Haibo Wu

PROVIDER: PXD081310 | Pride | 2026-10-19

REPOSITORIES: Pride