Stephen Buratowski PhD

The Buratowski Lab studies eukaryotic gene transcription, chromatin regulation, and RNA processing using a wide range of biochemical, molecular, and genetic techniques. The Buratowski Lab studies eukaryotic gene expression, concentrating on RNA polymerase II transcription and its coupling to RNA processing and chromatin modification. Much of our work centers on the C-terminal domain (CTD) of the Rpb1, the largest polymerase subunit. A dynamic pattern of differential phosphorylations act to mark polymerases at the initiation, early elongation, and late elongation stages. The mRNA capping enzyme and the histone H3K4 methyltransferase Set1 recognize the early phosphorylation pattern, while polyadenylation factors and the H3K36 methyltransferase Set2 binds the late CTD pattern. Much of this “CTD cycle” model is based on in vivo crosslinking, so a true understanding of the system dynamics will require more sophisticated methods. Towards this goal, we have adapted two technologies to our in vitro transcription system: time-resolved quantitative mass spectrometry and single-molecule TIRF microscopy. Combined with classic molecular genetics and genomics, these approaches are providing new insights into the kinetic and thermodynamic control of gene expression.