Protein partners of Kruppel-like factor 3

ABSTRACT: The aim of this study was to identify protein partners of the Kruppel-like factor 3 (KLF3) functional domain. We have previously shown that the non-DNA binding domain (the functional domain) of KLF3 play an important role in genomic localisation of this transcription factor (Burdach et al, 2014, Nucleic Acids Research, 42(1):276-89) and that the KLF3 functional domain is sufficient to direct an artificial zinc finger protein to new targets (Lim et al, 2016, Nucleic Acids Research, 44(7):3118-30). In this study we aimed to identify partner proteins of the KLF3 functional domain that might help to account for the role of this domain in genomic localisation. We had previously established HEK293 cell lines stably expressing either empty pMSCVpuro vector or pMSCVpuro-KLF3 functional domain (amino acids 1-262) with a C-terminal glycine-serine linker followed by a V5 epitope tag (Lim et al, 2016, Nucleic Acids Research, 44(7):3118-30). In this current study we performed two independent replicate experiments using the empty vector and KLF3 functional domain-V5 expressing HEK293 cells. Co-immunoprecipitation coupled with mass spectrometry was performed, using an antibody to the V5 tag, to identify endogenous HEK293 cell proteins that were bound by the KLF3 functional domain but not detected in the V5 co-immunoprecipitation samples from the empty vector cells. The mass spectrometry data was used to prioritise KLF3 functional domain partner proteins for further studies. In this experiment, WDR5 was identified as a partner protein of the KLF3 functional domain (in both independent replicates of the experiment) and the interaction of KLF3 and WDR5 was subsequently confirmed and the interaction interface of KLF3 that is bound by WDR5 mapped using co-immunoprecipitation experiments in COS cells.Sample description:- F002311 and F002312 are mass spec results from the empty transfected cell control cells- F002313 and F002314 are mass spec results from the cell stably expressing KLF3 functional domain

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell, Cell Culture

SUBMITTER: Kate Quinlan  

LAB HEAD: Kate Quinlan

PROVIDER: PXD033888 | Pride | 2025-09-23

REPOSITORIES: Pride