Doctor, Universitat Des Saarlandes (2011)
Diplom, Universitat Des Saarlandes (2004)
I am interested in the epigenetic reprogramming of DNA methylation during early mammalian preimplantation development. Early mammalian development is characterized by dramatic epigenetic changes. Upon fertilization of the oocyte with the sperm, the maternal and paternal genomes of the zygote are extensively reprogrammed to ensure the development of a totipotent potential. During this period of epigenetic reprogramming, DNA methylation (5-methyl-cytosine, 5mC) of paternal and maternal chromosomes is erased and reset during formation of the blastocyst. Interestingly, in mouse zygotes, the paternal genome becomes actively demethylated, as judged by immunofluorescence with antibodies against 5mC and bisulfite-sequencing data. Since the discovery of active DNA demethylation many scientists were trying to identify the putative “DNA demethylase” and a lot of candidate enzymes and pathways have been suggested and disproven. The identification of the enzymatic conversion of 5mC to 5-hydroxymethyl-cytosine (5hmC), 5-formyl-cytosine (5fC) and 5-carboxyl-cytosine (5caC) by Tet1-3 enzymes sheds new light on this process.
However, the analysis of epigenetic reprogramming in mammals is mainly focused on the mouse model and little is known about human embryonic development. Understanding the basic molecular mechanisms of human epigenetic reprogramming will impact human reproductive health and the generation of pluripotent stem cells
To ensure genome stability, pericentromeric regions are compacted in a dense heterochromatic structure through a combination of specific 'epigenetic' factors and modifications. A cascadal pathway is responsible for establishing pericentromeric chromatin involving chromatin modifiers and 'readers', such as H3K9 histone methyltransferases (Suv)39h and heterochromatin protein 1. Here we define how H3K64me3 on the lateral surface of the histone octamer integrates within the heterochromatinization cascade. Our data suggest that enrichment of H3K64me3 at pericentromeric chromatin foci is dependent on H3K9me3 but independent of a number of central factors such as heterochromatin protein 1, DNA methyltransferases and Suv4-20h histone methyltransferases. Our results support a model in which pericentromeric heterochromatin foci are formed along distinct pathways upon H3K9 trimethylation, involving H3K64me3 to potentially stabilize DNA-histone interactions, as well as sequential recruitment of repressive histone tail and DNA modifications. We hence suggest that multiple mechanisms ensure heterochromatin integrity at pericentromeres, with H3K64me3 as an important factor.
View details for DOI 10.1038/ncomms3233
View details for Web of Science ID 000323750400002
View details for PubMedID 23903902
The epigenomes of early mammalian embryos are extensively reprogrammed to acquire a totipotent developmental potential. A major initial event in this reprogramming is the active loss/demethylation of 5-methylcytosine (5mC) in the zygote. Here, we report on findings that link this active demethylation to molecular mechanisms. We detect 5-hydroxymethylcytosine (5hmC) as a novel modification in mouse, bovine and rabbit zygotes. On zygotic development 5hmC accumulates in the paternal pronucleus along with a reduction of 5mC. A knockdown of the 5hmC generating dioxygenase Tet3 simultaneously affects the patterns of 5hmC and 5mC in the paternal pronucleus. This finding links the loss of 5mC to its conversion into 5hmC. The maternal pronucleus seems to be largely protected against this mechanism by PGC7/Dppa3/Stella, as in PGC7 knockout zygotes 5mC also becomes accessible to oxidation into 5hmC. In summary, our data suggest an important role of 5hmC and Tet3 for DNA methylation reprogramming processes in the mammalian zygote.
View details for DOI 10.1038/ncomms1240
View details for Web of Science ID 000289982600031
View details for PubMedID 21407207
In mammalian zygotes, the 5-methyl-cytosine (5mC) content of paternal chromosomes is rapidly changed by a yet unknown but presumably active enzymatic mechanism. Here, we describe the developmental dynamics and parental asymmetries of DNA methylation in relation to the presence of DNA strand breaks, DNA repair markers and a precise timing of zygotic DNA replication. The analysis shows that distinct pre-replicative (active) and replicative (active and passive) phases of DNA demethylation can be observed. These phases of DNA demethylation are concomitant with the appearance of DNA strand breaks and DNA repair markers such as gammaH2A.X and PARP-1, respectively. The same correlations are found in cloned embryos obtained after somatic cell nuclear transfer. Together, the data suggest that (1) DNA-methylation reprogramming is more complex and extended as anticipated earlier and (2) the DNA demethylation, particularly the rapid loss of 5mC in paternal DNA, is likely to be linked to DNA repair mechanisms.
View details for DOI 10.1038/emboj.2010.80
View details for Web of Science ID 000278235100010
View details for PubMedID 20442707
Here, we summarize current knowledge about epigenetic reprogramming during mammalian preimplantation development, as well as the potential mechanisms driving these processes. We will particularly focus on changes taking place in the zygote, where the paternally derived DNA and chromatin undergo the most striking alterations, such as replacement of protamines by histones, histone modifications and active DNA demethylation. The putative mechanisms of active paternal DNA demethylation have been studied for over a decade, accumulating a lot of circumstantial evidence for enzymatic activities provided by the oocyte, protection of the maternal genome against such activities and possible involvement of DNA repair. We will discuss the various facets of dynamic epigenetic changes related to DNA methylation with an emphasis on the putative involvement of DNA repair in DNA demethylation.
View details for DOI 10.1387/ijdb.103206kl
View details for Web of Science ID 000291961200003
View details for PubMedID 21404179