Effect of nuclear architecture on translesion DNA synthesis

A growing body of evidence indicates that the nuclear architecture plays an important role in regulating gene expression, replication, and repair. We are performing an in-depth analysis of the role of nuclear architecture in TLS regulation in mammalian cells, including mapping genomic distribution of UV DNA damage along human chromosomes, mapping genomic sites of TLS activity in UV irradiated cells, and investigating the role of the nuclear sub-structures in TLS regulation. This is being done using the powerful techniques of chromatin-immuno-precipitation in combination with deep sequencing (ChIP-seq), and chromosome conformation capture (3C) in conjunction with deep sequencing (Hi-C). This work is done in collaboration with Dr. Amos Tanay’s group (Faculty of Mathematics and Computer Sciences). Additional approaches include localization by immuno-fluorescence microscopy, cell and nucleus fractionation, and a variety of cell biology and molecular biology techniques.