Craig M. Hart

 

Assistant Professor

Ph.D., Cornell University, 1989

Chromosome Organization, Chromatin Structure and Gene Expression

 

chart4@lsu.edu

 

BIOL 3090

 

Eukaryotic chromosomes are packaged into chromatin, whose repeating nucleosomal substructure is composed of DNA wrapped around histone proteins.  Nucleosomes interact with many other proteins to form dynamic, higher-order structures whose forms and functions are poorly understood.  My research is focused on the formidable challenge of understanding the role of chromatin dynamics and nuclear organization in essential nuclear processes such as gene expression.  My approach integrates the methods of biochemistry, genetics, and molecular and cell biology to understand the mechanisms of action of the class of chromosomal elements called chromatin domain insulator elements (also called boundary elements).  I am also interested in exploring the possibility that insulators can be developed into useful tools for reducing chromosomal position-effects on transgene expression levels in medical and research applications. 

 

Insulators define possible regulatory interactions within genomes.  They block enhancer-promoter communication, but only when located between the enhancer and promoter.  They could do this by acting as roadblocks that stop the propagation of active or repressed chromatin states.  To test this model, and to determine if different insulators are mechanistically similar, I am currently assaying the ability of various insulators to block the propagation of altered chromatin states in transgenic flies.  This is being done in collaboration with Dr. Pam Geyer of the University of Iowa.  I am also continuing to characterize the role of a protein I previously purified.  This protein, called BEAF (Boundary Element-Associated Factor), binds to sequences present in an abundant class of Drosophila insulators.  I purified another protein that can compete with BEAF for binding to a subset of BEAF-utilizing insulators, suggesting the activity of some insulators is regulated.  This protein is a transcription factor that regulates genes important for cell cycle progression, suggesting a link between BEAF activity and the cell cycle. 

 

Understanding how insulators limit communication between enhancers and promoters will address how enhancers find their target promoters, even when separated by tens of kilobases, without promiscuously interacting with other promoters.  The role played by insulators in regulating enhancer-promoter interactions suggests they might play a broader role in the organization of chromosomes into functionally separate domains.  Thus, understanding mechanisms governing insulator assembly and function should provide insight into the relationship between higher-order chromatin structure, nuclear organization, and gene regulation. 

 

 

Selected Publications

 

Hart, C.M., Cuvier, O., and Laemmli, U.K.  1999.  Evidence for an antagonistic relationship between the boundary element-associated factor BEAF and the transcription factor DREF.  Chromosoma 108:375-383. 

 

Cuvier, O., Hart, C.M., and Laemmli, U.K.  1998.  Identification of a class of chromatin boundary elements.  Mol. Cell. Biol. 18:7478-7486.

 

Hart, C.M. and Laemmli, U.K.  1998.  Facilitation of chromatin dynamics by SARs.  Curr. Opin. Genet. Dev. 8:519-525. 

 

Hart, C.M., Zhao, K., and Laemmli, U.K.  1997.  The scs’ boundary element: characterization of boundary element-associated factors.  Mol. Cell. Biol. 17:999-1009. 

 

Zhao, K., Hart, C.M., and Laemmli, U.K.  1995.  Vizualization of chromosomal domains with boundary element-associated factor BEAF-32.  Cell 81:879-889.