Periodic Reporting for period 1 - PRDM9Recomb (The Evolution of PRDM9 Binding and Genomic Localization of Meiotic Recombination Events)
Reporting period: 2015-05-01 to 2017-04-30
In another aspect of the project, we have mapped the chromatin landscape during meiosis. We have used several different markers of chromatin structure, including H3K4me3, H3K4me1, and H3K27ac. H3K4me3 is a chromatin modification that marks promoters and enhancers of genes and regions of open chromatin, but also marks sites of recombination events during meiosis, and is catalysed in part by PRDM9. As H3K4me1 and H3K27ac are also markers of promoters and enhancers, including these additional marks allows us to focus on the H3K4me3 marks that are independent and are not associated with gene regulatory elements, for these are the regions that are most likely to be associated with PRDM9 and meiotic recombination. Furthermore, we have similarly mapped these chromatin markers in other somatic tissues, and have found that there is much more H3K4me3 that is unique to the testes and is not associated with gene regularity elements, and that this H3K4me3 is likely to be due to the activity of PRDM9. We will further investigate the evolution of these chromatin marks in different mammals where PRMD9 is not playing a role during meiotic recombination.
Furthermore, we wanted to investigate the role that retrotransposons may being playing in shaping meiotic recombination and genome evolution. During meiosis, the structure of the chromatin changes and many transposons that are normally suppressed become activated. This has severe implications for genome evolution, as any new insertions and mutations that get induced will be inherited by the next generation. Furthermore, as transposon activity causes breaks in the DNA, we were curious if there was an interplay between meiotic recombination and transposon activity. As LINE-1 is the only autonomous transposon in humans, and all active retrotransposons use LINE-1's machinery to transpose, we wanted to investigate the proteins encoded by LINE-1 to see if there was any association with meiotic recombinant hotspots. We carried out immunoprecipitation agains LINE-1 proteins to look for any enrichment in recombination sequences in the interacting nucleotides.
Progress on this project is ongoing. In particular we are investigating the relationship between the evolution of chromatin structure and organisation, and the evolution of meiotic recombination and PRDM9 association with chromatin. In the follow-up work that I will be carrying out now, we are expanding our study to look across 10 different species of mammals, rather than restricting the study only to mouse strains. In this study we will be able to look at the impact of PRDM9's activity on recombination landscape and also on chromatin structure and other aspects of genome function, such as gene expression. This study will give us insight into the interplay between genome evolution, chromatin structure, and the meiotic recombination landscape in mice and across mammals.