introductory-analysis-and-validation-of-cutrun-sequencing-data

Introductory Analysis and Validation of CUT&#38;RUN Sequencing Data

The CUT&amp;RUN technique facilitates detection of protein-DNA interactions across the genome. Typical applications of CUT&amp;RUN include profiling ...

University of Texas MD Anderson Cancer Center

The SWI/SNF ATP-dependent chromatin remodeler is a master regulator of the epigenome, controlling pluripotency and differentiation. Towards the C-terminus of the catalytic subunit of SWI/SNF is a motif called the AT-hook that is evolutionary conserved. The AT-hook is present in many chromatin modifiers and generally thought to help anchor them to DNA. We observe however that the AT-hook regulates the intrinsic DNA-stimulated ATPase activity aside from promoting SWI/SNF recruitment to DNA or nucleosomes by increasing the reaction velocity a factor of 13 with no accompanying change in substrate affinity (K). The changes in ATP hydrolysis causes an equivalent change in nucleosome movement, confirming they are tightly coupled. The catalytic subunit's AT-hook is required in vivo for SWI/SNF remodeling activity in yeast and mouse embryonic stem cells. The AT-hook in SWI/SNF is required for transcription regulation and activation of stage-specific enhancers critical in cell lineage priming. Similarly, growth assays suggest the AT-hook is required in yeast SWI/SNF for activation of genes involved in amino acid biosynthesis and metabolizing ethanol. Our findings highlight the importance of studying SWI/SNF attenuation versus eliminating the catalytic subunit or completely shutting down its enzymatic activity.

Junwoo Lee

Department of Epigenetics and Molecular Carcinogenesis

University of Texas MD Anderson Cancer Center

Introductory Analysis and Validation of CUT&RUN Sequencing Data

Junwoo Lee

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Epigenetics and Molecular Carcinogenesis

University of Texas MD Anderson Cancer Center

Introductory Analysis and Validation of CUT&RUN Sequencing Data

Department of Epigenetics and Molecular Carcinogenesis