Streamlined histone-based fluorescence lifetime imaging microscopy (FLIM) for studying chromatin organisation

Changes in chromatin structure are key determinants of genomic responses. Thus, methods that enable such measurements are instrumental for investigating genome regulation and function. Here, we report further developments and validation of a streamlined method of histone-based fluorescence lifetime imaging microscopy (FLIM) that robustly detects chromatin compaction states in fixed and live cells, in 2D and 3D. We present a quality-controlled and detailed method that is simpler and faster than previous methods, and uses FLIMfit open-source software. We demonstrate the versatility of this chromatin FLIM through its combination with immunofluorescence and implementation in immortalised and primary cells. We applied this method to investigate the regulation of chromatin organisation after genotoxic stress and provide new insights into the role of ATM in controlling chromatin structure independently of DNA damage. Collectively, we present an adaptable chromatin FLIM method for examining chromatin structure and establish its utility in mammalian cells

Correction: LGR5 regulates pro-survival MEK/ERK and proliferative Wnt/β-catenin signalling in neuroblastoma

A Correction on: LGR5 regulates pro-survival MEK/ERK and proliferative Wnt/β-catenin signalling in neuroblastoma Gabriella Cunha Vieira, S. Chockalingam, Zsombor Melegh, Alexander Greenhough, Sally Malik, Marianna Szemes, Ji Hyun Park, Abderrahmane Kaidi, Li Zhou, Daniel Catchpoole, Rhys Morgan, David O. Bates, Peter J. Gabb and Karim Malik Original article: Oncotarget. 2015; 6:40053-67. DOI: 10.18632/oncotarget.5548. The originally Figure 5 contains duplicate total-ERK panels. The proper Figure 5 is attached. The authors sincerely apologize for this error

The role of hypoxia in colorectal tumorigenesis

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

KAT5 tyrosine phosphorylation couples chromatin sensing to ATM signalling

Detecting genomic changes represents a critical step in cellular responses to DNA damage. Here, we show that tyrosine phosphorylation of the protein acetyltransferase KAT5 (Tip60) increases in response to DNA damage in a manner that promotes KAT5 binding to the histone mark H3K9me3. This in turn triggers KAT5-mediated acetylation of the ATM kinase, promoting DNA-damage checkpoint activation and cell survival. We also establish that chromatin alterations per se can enhance KAT5 tyrosine phosphorylation and ATM-dependent signaling, and identify the proto-oncogene c-Abl as mediating this modification. These findings define KAT5 as a key sensor for genomic and chromatin perturbations, and highlight a prime role for c-Abl in such events