Proteomic analysis identifies ZMYM2 as endogenous binding partner of TBX18 protein in 293 and A549 cells. Issue 1 (14th January 2022)

• Record Type: Journal Article
• Authors: Lüdtke, Timo H.-W.; Kleppa, Marc-Jens; Rivera-Reyes, Reginaldo; Qasrawi, Fairouz; Connaughton, Dervla M.; Shril, Shirlee; Hildebrandt, Friedhelm; Kispert, Andreas
• Is Part Of: Biochemical journal. Volume 479:Issue 1(2022)
• Page Start: 91
• Page End: 109
• Publication Date: 2022-01-14
• Subjects: protein–protein interactions -- proteomics -- proximity ligation assay -- transcription factor; Biochemistry -- Periodicals; 572
• DOI: 10.1042/BCJ20210642
• Languages: English
• ISSNs: 0264-6021

Abstract

Abstract : The TBX18 transcription factor regulates patterning and differentiation programs in the primordia of many organs yet the molecular complexes in which TBX18 resides to exert its crucial transcriptional function in these embryonic contexts have remained elusive. Here, we used 293 and A549 cells as an accessible cell source to search for endogenous protein interaction partners of TBX18 by an unbiased proteomic approach. We tagged endogenous TBX18 by CRISPR/Cas9 targeted genome editing with a triple FLAG peptide, and identified by anti-FLAG affinity purification and subsequent LC–MS analysis the ZMYM2 protein to be statistically enriched together with TBX18 in both 293 and A549 nuclear extracts. Using a variety of assays, we confirmed the binding of TBX18 to ZMYM2, a component of the CoREST transcriptional corepressor complex. Tbx18 is coexpressed with Zmym2 in the mesenchymal compartment of the developing ureter of the mouse, and mutations in TBX18 and in ZMYM2 were recently linked to congenital anomalies in the kidney and urinary tract (CAKUT) in line with a possible in vivo relevance of TBX18–ZMYM2 protein interaction in ureter development.