- Senior Investigator, Nuclear Organization and Gene Expression Section, Laboratory of Cellular and Developmental Biology, NIDDK, NIH, 2014-Present
- Investigator, Laboratory of Cellular and Developmental Biology NIDDK, NIH, 2006-2014
- Postdoctoral Fellow, Laboratory of Victor G. Corces, Johns Hopkins University, NIDDK, NIH, 2003-2006
- Ph.D., Laboratory of Pamela A. Silver, Harvard Medical School, 1997-2002
The work performed in my laboratory will help determine how the 3-dimensional organization of DNA within each cell affects cellular function and identity throughout development.
Importance of chromatin insulators
It has become increasingly apparent that proper control of gene expression requires complex organization of DNA at the level of chromatin. Chromatin insulators are DNA-protein complexes that influence gene expression by establishing chromatin domains subject to distinct transcriptional controls, likely through alteration of their spatial organization. Insulators enforce the strict specific and temporal expression of loci with complex enhancer and/or promoter configuration. Examples include metazoan Hox genes, master regulators of body segmentation, and the vertebrate beta-globin locus, which changes in expression during erythroid development. Loss of insulator activity can result in substantial positive or negative changes in gene expression, culminating in disease, defects in development, and/or lethality. For example, deletion of insulator binding sites at the H19/IGF2 imprinting center have been implicated in Beckwith-Wiedemann syndrome and Wilms’ Tumor. Moreover, recent studies have shown that loss of insulator activity in IDH1 mutant gliomas and T cell acute lymphoblastic leukemias leads to disruption of boundaries between chromatin domains and subsequent oncogene activation.
The gypsy chromatin insulator
We primarily utilize the biochemically and genetically tractable model system Drosophila, which harbors the largest diversity of known chromatin insulator complexes. Defined by the specific binding of the Su(Hw) zinc finger DNA-binding protein, gypsy insulator complexes tend to associate with gene-poor, transcriptionally inert regions of the genome. Within the nucleus, gypsy insulator complexes concentrate at approximately 200nm diameter ovoid structures termed insulator bodies, which are tethered stably to the nuclear matrix. The proper localization of insulator bodies is highly correlated with gypsy chromatin insulator function, but their precise function and spatial relationship with respect to the genome is not well understood. We are investigating the cofactors required for and the ultrastructure of insulator bodies within the surrounding chromatin environment.
Tissue-specific regulation of insulator activity
We recently identified two novel, tissue-specific negative regulators of gypsy insulator function that affect both enhancer blocking and barrier activities. Shep can bind directly to Su(Hw) as well as another core component of the gypsy insulator complex, potentially competing with inter- or intra-insulator complex interactions and thereby neutralizing insulator activity. Shep is required for neuronal remodeling during development and is highly enriched in the CNS, perhaps serving to negatively regulate insulator function in these cell types to promote CNS-specific gene expression programs. Shep harbors two highly conserved RNA recognition motifs (RRMs), and genetic evidence points to a functional relationship between its RNA-binding capability and insulator function. In contrast, Rump, which contains 3 RRMs, antagonizes gypsy insulator activity in tissues outside of the CNS.
RNA-dependent insulator function
Using RNA immunoprecipitation followed by deep sequencing (RIP-seq), we made the striking finding that certain mRNAs, including that encoding Su(Hw) itself, associate stably with gypsy insulator complexes. Expression of untranslatable versions of these mRNAs alters insulator body localization and promotes insulator activity. We speculate that these, and possibly other mRNAs, also harbor a noncoding function, such as acting as a scaffold for insulator complexes at specific subnuclear locations. We continue to delve into the mechanisms by which these mRNAs function and the roles of associated RNA-binding proteins in insulator regulation.
(This work was highlighted in Editors' Choice, Riddihough G. Noncoding mRNAs. Science, (341)6149: 938, 2013.) Link: http://www.sciencemag.org/content/341/6149/twil.full
Need for Further Study
Better understanding of the mechanisms of chromatin insulator function, including the identification of novel interactors and regulatory steps, are needed to move the field forward.
- Argonaute2 and LaminB modulate gene expression by controlling chromatin topology.
- Nazer E, Dale RK, Chinen M, Radmanesh B, Lei EP.
- PLoS Genet (2018 Mar) 14:e1007276. Abstract/Full Text
- Shep regulates Drosophila neuronal remodeling by controlling transcription of its chromatin targets.
- Chen D, Dale RK, Lei EP.
- Development (2018 Jan 3) 145. Abstract/Full Text
Research in Plain Language
Each cell within the body contains the identical genetic information, but the complement of genes that are active at any given time is a critical factor in determining the type of cell (i.e. kidney, lung, heart) and its specific function within a particular tissue. All of the genetic information is contained within the DNA, and if the DNA were stretched end to end, it would extend 2 meters. Therefore, the DNA must be tightly packaged in order to fit within the cell. At the same time, the genes within the DNA must be kept accessible to the cellular machinery in order to be read as the genetic blueprint to produce RNA and proteins, the actual workhorses of the cell. The DNA is wrapped around a variety of proteins that help compact and organize its overall structure. One such complex of DNA-bound protein is called a chromatin insulator. With respect to the genetic blueprint, chromatin insulators function similarly to punctuation within a paragraph, in order to help subdivide otherwise incoherent strings of words into sentences and to logically connect related phrases.
My laboratory studies how chromatin insulators function and how they help control which genes are active. Studying how chromatin insulators work helps improve our understanding of how cells develop and give rise to different tissues. When chromatin insulators malfunction, disease can result. In fact, a single cell cannot survive without the activity of chromatin insulators.
Since chromatin insulators exist and function similarly in most multicellular animal organisms, we use the relatively simple fruit fly Drosophila as a model organism. Working with Drosophila is advantageous to doing experiments in mammals because of their much faster generation time and ease of use in genetic experiments. The Drosophila system is also extremely powerful for biochemistry and cell biology techniques, and the Drosophila genome has the largest diversity of known chromatin insulator proteins.