Promiscuous DNA-binding of a mutant zinc finger protein corrupts the transcriptome and diminishes cell viability

The rules of engagement between zinc finger transcription factors and DNA have been partly defined by in vitro DNA-binding and structural studies, but less is known about how these rules apply in vivo. Here we demonstrate how a missense point mutation in the second zinc finger of Krüppel-like factor-1 (KLF1) leads to degenerate DNA-binding specificity in vivo, resulting in ectopic transcription and semi-dominant neonatal anemia. We employed RNA-seq, ChIP-seq and 4sU-RNA-seq to identify the direct transcriptional consequences of aberrant DNA-binding events genome wide. Funding: National Health and Medical Research Council [APP1082429 to A.C.P.]; United States Public Health Service [R01 GM103544 to T.B., R01 DK100692 to L.L.P. and R01 DK46865 to J.J.B.]; Victorian Government Operational Infrastructure Support Scheme [St Vincent's Institute of Medical Research to M.W.P]. Funding for open access charge: Mater Research - UQ [APP1082429]. mRNA sequencing Klf1Nan/+ and wild type fetal liver (E14.5) identified a requirement for in vitro studies. ChIP-seq and 4sU-RNA-seq were performed on replicate immortalized cell lines transfected with tamoxifen inducible protein fusions of KLF1-ER or Nan-KLF1-ER.