5EGB: Human Prdm9 Allele-a Znf Domain With Associated Recombination Hotspot Dna Sequence Ii

Citation:
Abstract
The multidomain zinc finger (ZnF) protein PRDM9 (PRD1-BF1-RIZ1 homologous domain-containing 9) is thought to influence the locations of recombination hot spots during meiosis by sequence-specific DNA binding and trimethylation of histone H3 Lys4. The most common variant of human PRDM9, allele A (hPRDM9A), recognizes the consensus sequence 5'-NCCNCCNTNNCCNCN-3'. We cocrystallized ZnF8-12 of hPRDM9A with an oligonucleotide representing a known hot spot sequence and report the structure here. ZnF12 was not visible, but ZnF8-11, like other ZnF arrays, follows the right-handed twist of the DNA, with the alpha helices occupying the major groove. Each alpha helix makes hydrogen-bond (H-bond) contacts with up to four adjacent bases, most of which are purines of the complementary DNA strand. The consensus C:G base pairs H-bond with conserved His or Arg residues in ZnF8, ZnF9, and ZnF11, and the consensus T:A base pair H-bonds with an Asn that replaces His in ZnF10. Most of the variable base pairs (N) also engage in H bonds with the protein. These interactions appear to compensate to some extent for changes from the consensus sequence, implying an adaptability of PRDM9 to sequence variations. We investigated the binding of various alleles of hPRDM9 to different hot spot sequences. Allele C was found to bind a C-specific hot spot with higher affinity than allele A bound A-specific hot spots, perhaps explaining why the former is dominant in A/C heterozygotes. Allele L13 displayed higher affinity for several A-specific sequences, allele L9/L24 displayed lower affinity, and allele L20 displayed an altered sequence preference. These differences can be rationalized structurally and might contribute to the variation observed in the locations and activities of meiotic recombination hot spots.
PDB ID: 5EGBDownload
MMDB ID: 136597
PDB Deposition Date: 2015/10/26
Updated in MMDB: 2017/10
Experimental Method:
x-ray diffraction
Resolution: 1.98  Å
Source Organism:
Similar Structures:
Biological Unit for 5EGB: trimeric; determined by author and by software (PISA)
Molecular Components in 5EGB
Label Count Molecule
Protein (1 molecule)
1
Histone-lysine N-methyltransferase Prdm9(Gene symbol: PRDM9)
Molecule annotation
Nucleotides(2 molecules)
1
DNA (5'- D(*ap*cp*cp*ap*cp*gp*tp*gp*gp*cp*tp*ap*gp*gp*gp*ap*gp*gp*cp*cp*t)- 3')
Molecule annotation
1
DNA (5'- D(*tp*ap*gp*gp*cp*cp*tp*cp*cp*cp*tp*ap*gp*cp*cp*ap*cp*gp*tp*gp*g)- 3')
Molecule annotation
Chemicals (4 molecules)
1
4
* Click molecule labels to explore molecular sequence information.

Citing MMDB
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