RT Journal Article
SR Electronic
T1 PRDM9 forms a trimer by interactions within the zinc finger array
JF Life Science Alliance
JO Life Sci. Alliance
FD Life Science Alliance LLC
SP e201800291
DO 10.26508/lsa.201800291
VO 2
IS 4
A1 Schwarz, Theresa
A1 Striedner, Yasmin
A1 Horner, Andreas
A1 Haase, Karin
A1 Kemptner, Jasmin
A1 Zeppezauer, Nicole
A1 Hermann, Philipp
A1 Tiemann-Boege, Irene
YR 2019
UL http://www.life-science-alliance.org/content/2/4/e201800291.abstract
AB 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 in vitro 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.