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 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 anemia in the Nan mouse model. We employed ChIP-seq and 4sU-RNA-seq to identify aberrant DNA-binding events genome wide and ectopic transcriptional consequences of this binding. We confirmed novel sequence specificity of the mutant recombinant zinc finger domain by performing biophysical measurements of in vitro DNA-binding affinity. Together, these results shed new light on the mechanisms by which missense mutations in DNA-binding domains of transcription factors can lead to autosomal dominant diseases.
Cancer Genomics Group, Mater Research Institute - University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia.
Kevin R
KR
Gillinder
Cancer Genomics Group, Mater Research Institute - University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia.
Melissa D
MD
Ilsley
The Jackson Laboratory, Bar Harbor, ME, USA.
Danitza
D
Nébor
Department of Oncological Sciences, Mount Sinai School of Medicine, New York, NY, USA.
Ravi
R
Sachidanandam
Institute for Molecular Bioscience, University of Queensland, Brisbane, QLD, Australia.
Mathieu
M
Lajoie
Cancer Genomics Group, Mater Research Institute - University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia.
Graham W
GW
Magor
Cancer Genomics Group, Mater Research Institute - University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia.
Michael R
MR
Tallack
Department of Pharmacology, School of Medicine, University of Nevada, Reno, NV, USA.
Timothy
T
Bailey
School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD, Australia.
Michael J
MJ
Landsberg
School of Life and Environmental Sciences, The University of Sydney, NSW, Australia.
Joel P
JP
Mackay
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Melbourne, VIC, Australia.|ACRF Rational Drug Discovery Centre, St. Vincent's Institute of Medical Research, Melbourne, VIC, Australia.
Michael W
MW
Parker
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Melbourne, VIC, Australia.
Luke A
LA
Miles
The Jackson Laboratory, Bar Harbor, ME, USA.
Joel H
JH
Graber
The Jackson Laboratory, Bar Harbor, ME, USA.
Luanne L
LL
Peters
Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine, New York, NY, USA.
James J
JJ
Bieker
Cancer Genomics Group, Mater Research Institute - University of Queensland, Translational Research Institute, Woolloongabba, QLD, Australia.|Princess Alexandra Hospital, Brisbane, QLD 4102, Australia.
Andrew C
AC
Perkins
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D041905:Erythroid Cells / Q000378:metabolism; D004920:Erythropoiesis / Q000235:genetics; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008957:Models, Genetic; D008958:Models, Molecular; D050505:Mutant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020125:Mutation, Missense; D011485:Protein Binding; D059467:Transcriptome / Q000235:genetics*; D016335:Zinc Fingers / Q000235:genetics*
D051741:Kruppel-Like Transcription Factors; D050505:Mutant Proteins; C080774:erythroid Kruppel-like factor; D004247:DNA