PRDM9 forms a trimer by interactions within the zinc finger array

PRDM9 is a trans-acting factor directing meiotic recombination to specific DNA-binding sites by its zinc finger (ZnF) array. It was suggested that PRDM9 is a multimer; however, we do not know the stoichiometry or the components inducing PRDM9 multimerization. In this work, we used binding studies an...

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Bibliographic Details
Published in:Life science alliance 2019-08, Vol.2 (4), p.e201800291
Main Authors: Schwarz, Theresa, Striedner, Yasmin, Horner, Andreas, Haase, Karin, Kemptner, Jasmin, Zeppezauer, Nicole, Hermann, Philipp, Tiemann-Boege, Irene
Format: Article
Language:English
Subjects:
Animals
Binding Sites
DNA - chemistry
DNA - metabolism
Electrophoretic Mobility Shift Assay
Histone-Lysine N-Methyltransferase - chemistry
Histone-Lysine N-Methyltransferase - metabolism
Homologous Recombination
Mass Spectrometry
Meiosis
Mice
Models, Molecular
Protein Multimerization
Solubility
Zinc Fingers
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Summary:PRDM9 is a trans-acting factor directing meiotic recombination to specific DNA-binding sites by its zinc finger (ZnF) array. It was suggested that PRDM9 is a multimer; however, we do not know the stoichiometry or the components inducing PRDM9 multimerization. In this work, we used binding studies and characterized with electrophoretic mobility shift assays, mass spectrometry, and fluorescence correlation spectroscopy the stoichiometry of the PRDM9 multimer of two different murine PRDM9 alleles carrying different tags and domains produced with different expression systems. Based on the migration distance of the PRDM9-DNA complex, we show that PRDM9 forms a trimer. Moreover, this stoichiometry is adapted already by the free, soluble protein with little exchange between protein monomers. The variable ZnF array of PRDM9 is sufficient for multimerization, and at least five ZnFs form already a functional trimer. Finally, we also show that only one ZnF array within the PRDM9 oligomer binds to the DNA, whereas the remaining two ZnF arrays likely maintain the trimer by ZnF-ZnF interactions.
ISSN:2575-1077
2575-1077
DOI:10.26508/lsa.201800291