The long-term objectives of the proposed project are to elucidate the mechanism of action and in vivo function of Mot1, an essential yeast transcriptional regulator that can activate or repress transcription. Mot1 is a member of a large family of evolutionarily conserved nuclear ATPases (the Snf2/Swi2 family) involved in transcription, DNA repair, and recombination. Defects in human Snf2/Swi2-related protein complexes are known to contribute to certain pediatric cancers, Cockayne's Syndrome, a-thalassemia, and the most common form of X-linked mental retardation. Despite the ubiquitous occurrence of Snf2/Swi2 family members, the molecular mechanisms of action of these proteins are not understood in detail, nor is it understood what roles many of these proteins play in vivo. Mot1 's ATPase activity is required to activate or repress transcription of specific genes in vivo. Consistent with its role as a repressor, Mot1 can dissociate TATA-binding protein (TBP)-DNA complexes in an ATP-dependent reaction. Motl's mechanism of ATP-dependent transcriptional activation is unknown. Biochemical, molecular biological, and genetic approaches will be used to define how Mot1 regulates transcription in vivo and how the Mot1 response of particular promoters is determined. These approaches will also be used to understand how ATP hydrolysis by Mot1 drives TBP/DNA disruption. The proposed analysis of Mot1 function will lead to a better understanding of the role of Motl in transcriptional control as well as a better understanding of the functions of Snf2/Swi2 family members in general.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM055763-08
Application #
6729055
Study Section
Cell Development and Function Integrated Review Group (CDF)
Program Officer
Tompkins, Laurie
Project Start
1997-05-01
Project End
2006-04-30
Budget Start
2004-05-01
Budget End
2005-04-30
Support Year
8
Fiscal Year
2004
Total Cost
$286,971
Indirect Cost
Name
University of Virginia
Department
Biochemistry
Type
Schools of Medicine
DUNS #
065391526
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
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Hoffman, Elizabeth A; Frey, Brian L; Smith, Lloyd M et al. (2015) Formaldehyde crosslinking: a tool for the study of chromatin complexes. J Biol Chem 290:26404-11
Viswanathan, Ramya; Hoffman, Elizabeth A; Shetty, Savera J et al. (2014) Analysis of chromatin binding dynamics using the crosslinking kinetics (CLK) method. Methods 70:97-107
Poorey, Kunal; Viswanathan, Ramya; Carver, Melissa N et al. (2013) Measuring chromatin interaction dynamics on the second time scale at single-copy genes. Science 342:369-72

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