MAMMALIAN mRNA TRANSCRIPTION AND ITS REGULATION:
Controlling gene expression is essential to growth, development, and sustained life. A critical control point for regulating gene expression is at the level of transcription. The proper regulation of transcription is essential for maintaining normal pathways of cell growth and differentiation, thereby avoiding the rampant cell proliferation observed in tumors. Transcription of protein encoding genes in eukaryotes is orchestrated by a host of protein factors, including RNA polymerase II, general transcription factors (TFIIA, TFIIB, TFIID, TFIIE, TFIIF, and TFIIH), coactivators, chromatin remodeling factors, gene-specific transcriptional regulatory proteins (activators and repressors), as well as non-coding RNAs. The underlying goal of our work is to uncover molecular mechanisms governing mammalian RNA polymerase II transcription and its regulation. To this end, we use a combination of biochemistry, molecular biology, molecular genetics, cell-based assays, genomics, and single molecule techniques to investigate mechanisms of mammalian transcriptional regulation.
Regulation of RNA Polymerase II by non-coding RNAs
All cells respond to stress, and do so in part by altering gene expression. When eukaryotic cells are subjected to heat shock, general RNA polymerase II transcription decreases at the same time as transcription of a set of heat shock specific genes increases. Two non-coding RNAs (mouse B2 RNA and human Alu RNA) are transcriptionally upregulated upon heat shock. We have found that these ncRNAs bind RNA polymerase II with high affinity (low nM) and block the formation of functional initiation complexes in vitro..... -more-
Regulation of Human mRNA Transcription by NFAT and AP-1 Proteins
The mammalian immune system represents a unique model for studying the importance of transcriptional regulation in governing cell growth and differentiation. Interleukin-2 is a cytokine that acts as an autocrine growth factor promoting the proliferation and development of T cells during the immune response to bacterial and viral infection, as well as tumorigenesis...... -more-
Single Molecule Approaches to Understanding Protein-Nucleic Acid Binding and the RNA polymerase II Transcription Reaction
The goal of this line of research is to investigate the assembly mechanism, dynamics, and heterogeneity of human transcription factor/DNA complexes, and how these parameters contribute to transcriptional control. Single-molecule studies have emerged as essential contributors to understanding the dynamic behavior, conformational states, and heterogeneity of biological complexes, thus allowing unprecedented insight into their function....... -more-