Characterization of the DNA Binding Activity of the ZFY Zinc Finger Domain

Abstract

The ZFY protein is a member of one of the most interesting classes of polydactyl zinc finger proteins. It has a domain of 13 tandem zinc fingers that is organized with an internal repeat of odd-even finger pairs. It has been proposed that each finger pair interacts with six base pairs within a turn of the double helix, the downstream linker crossing the minor groove to place the next finger pair on the following turn of the DNA. Yet putative binding sites for the full-length ZFY protein appear to consist of a six-base AGGCCY consensus sequence that is present in one or two copies. In this study the equilibrium binding of two ZFY-derived zinc finger peptides to 4R DNA with tandem copies of the consensus sequence was investigated. The ZFY5 peptide contains fingers 5-13, including four odd-even finger pairs, and the ZFY11 peptide contains fingers 11-13 and has one odd-even finger pair. Both peptides bound to 4R DNA with equal affinities, forming a bimolecular complex that is mediated by the downstream AGGCCY motif. The additional odd-even finger pairs in ZFY5 made no measurable difference in the mechanism of DNA binding compared to ZFY11. The effects on the DNA-protein interaction of mutations in the 4R DNA and in the key alpha-helical residues of fingers 11-13 indicate that the binding of ZFY to DNA is mediated by the interaction of the GGCC core base pairs with fingers 12 and 13. These results demonstrate that the even-odd repeats in the ZFY zinc finger domain do not make significant contributions to DNA binding.

The expression of Y-linked Zfy2 in XY mouse oocytes leads to frequent meiosis 2 defects, a high incidence of subsequent early cleavage stage arrest and infertility

“…The full-length proteins encoded by Zfy1 and Zfy2 are predicted to be zinc-finger transcription factors that bind the same DNA target sequences (Grants et al, 2010; Taylor-Harris et al, 1995); this suggests that Zfy1 may contribute to the impaired fertility of XY females. We therefore introduced into XO females either a single copy of a Y genomic BAC containing Zfy1 and Uba1y that had inserted on the X chromosome ( Zfy1 lo ) or 13-14 copies of the same BAC inserted on an autosome ( Zfy1 hi ) (Royo et al, 2010).…”

Quantitative analysis of ZFY and CTCF reveals dependent recognition of tandem zinc finger proteins

“…2B). Taylor-Harris et al [40] performed SELEX on mZFY1 and found it preferentially binds to sequences containing GGCCT and Grants et al [41] showed that fingers 11-13 were sufficient to recognize a motif of RGGCCT, both of which are consistent with the model from zinc finger motif prediction algorithms ( Fig. 2B) [29].…”

“…Human ZFY was used as reference so for other proteins only those different portions were shown; B) Motif prediction for human ZFY [29], in which positions 1-39 are recognized by Finger 13 to 1 and aligned with finger pattern in Fig. 1A; C) Motif from mouse ZFX PBM data [41]; D) Motif from mouse ZFX Chip-seq data [42]; E) Human ZFY motif from Affinity-seq data; F) Mouse ZFY1 motif from Affinity-seq data. MEME was used as the default motif-finding program.…”

Mouse Y-Encoded Transcription Factor Zfy2 Is Essential for Sperm Head Remodelling and Sperm Tail Development

“…The transcription factors encoded by Zfy1 and Zfy2 , together with their X-linked homologue Zfx , are predicted to bind the same DNA sequence [ 26 – 28 ]. In most eutherian mammals Zfx and Zfy are widely expressed, and Zfx is exempt from X dosage compensation, suggesting a constraining dosage requirement in somatic tissues [ 29 , 30 ].…”