Abstract
Myelin transcription factor 1 (MyT1/NZF2), a member of the neural zinc-finger (NZF) protein family, is a transcription factor that plays a central role in the developing central nervous system. It has also recently been shown that, in combination with two other transcription factors, the highly similar paralog MyT1L is able to direct the differentiation of murine and human stem cells into functional neurons. MyT1 contains seven zinc fingers (ZFs) that are highly conserved throughout the protein and throughout the NZF family. We recently presented a model for the interaction of the fifth ZF of MyT1 with a DNA sequence derived from the promoter of the retinoic acid receptor (RARE) gene. Here, we have used NMR spectroscopy, in combination with surface plasmon resonance and data-driven molecular docking, to delineate the mechanism of DNA binding for double ZF polypeptides derived from MyT1. Our data indicate that a two-ZF unit interacts with the major groove of the entire RARE motif and that both fingers bind in an identical manner and with overall two-fold rotational symmetry, consistent with the palindromic nature of the target DNA. Several key residues located in one of the irregular loops of the ZFs are utilized to achieve specific binding. Analysis of the human and mouse genomes based on our structural data reveals three putative MyT1 target genes involved in neuronal development.
Highlights
Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA
We showed that the F4F5RARE complex forms with a 1:1 stoichiometry
Differences in the DNA Binding Mode between MyT1 Single and Double Fingers—In this study, we have used a combination of NMR and SPR data to calculate a structural model of a MyT1 double zinc fingers (ZFs) domain bound to DNA
Summary
Myelin transcription factor 1 (MyT1) contains seven similar zinc finger domains that bind DNA . It has recently been shown that, in combination with two other transcription factors, the highly similar paralog MyT1L is able to direct the differentiation of murine and human stem cells into functional neurons. MyT1 was first discovered through its ability to bind to sites in the proteolipid protein promoter [2] This protein [3] plays a major role in the structure and compaction of the myelin sheath that is located around the axons of the central nervous system. Our NMR data show that this binding mode is conserved among other MyT1 double ZF constructs, indicating that the full-length protein might be able to recognize two or three distinct AAGTT sites in vivo
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