chromatin structure regulates gene conversion染色质结构调节基因转换.pdfVIP

PLoS BIOLOGY Chromatin Structure Regulates Gene Conversion 1 1 2 1 1 1,2* W. Jason Cummings , Munehisa Yabuki , Ellen C. Ordinario , David W. Bednarski , Simon Quay , Nancy Maizels 1 Department of Immunology, University of Washington School of Medicine, Seattle, Washington, United States of America, 2 Department of Biochemistry, University of Washington School of Medicine, Seattle, Washington, United States of America Homology-directed repair is a powerful mechanism for maintaining and altering genomic structure. We asked how chromatin structure contributes to the use of homologous sequences as donors for repair using the chicken B cell line DT40 as a model. In DT40, immunoglobulin genes undergo regulated sequence diversification by gene conversion templated by pseudogene donors. We found that the immunoglobulin Vk pseudogene array is characterized by histone modifications associated with active chromatin. We directly demonstrated the importance of chromatin structure for gene conversion, using a regulatable experimental system in which the heterochromatin protein HP1 (Drosophila melanogaster Su[var]205), expressed as a fusion to Escherichia coli lactose repressor, is tethered to polymerized lactose operators integrated within the pseudo-Vk donor array. Tethered HP1 diminished histone acetylation within the pseudo-Vk array, and altered the outcome of Vk diversification, so that nontemplated mutations rather than templated mutations predominated. Thus, chromatin structure regulates homology-directed repair. These results suggest that histone modifications may contribute to maintaining