Molecular Studies Of Protein-DNA Interactions

Hsieh, Peggy

Abstract

Using a variety of biochemical, structural, and molecular and cell biological approaches, we are studying the molecular mechanisms underlying the diverse array of cellular functions involving DNA repair proteins. Surface plasmon resonance studies allow us to monitor in real-time key protein-protein and protein-DNA interactions formed in vitro under a variety of conditions to ask about complex stability and to characterize various aspects of complex formation utilizing both wild type and mutant proteins. Fluorescently tagged proteins and immunofluorescence microscopy are being used to study protein-protein interactions as well as traffic within different cell compartments. Modulation of the levels of repair proteins, post-translational modifications, and subcellular localization are being examined in response to a variety of genotoxic stresses. Single molecule studies using atomic force microscopy allow us to visualize multi-protein complexes bound to DNA lesions. These studies can help shed light on specific DNA repair pathways as well as general cellular responses to DNA damage that may be relevant to cancer and aging.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
• Type: Intramural Research (Z01)
• Project #: 1Z01DK052015-18
• Application #: 7593637
• Support Year: 18
• Fiscal Year: 2007
• Total Cost: $410,774

Institution

• Name: National Institute of Diabetes and Digestive and Kidney Diseases
• Country: United States

Publications

Li, Zhongdao; Pearlman, Alexander H; Hsieh, Peggy (2016) DNA mismatch repair and the DNA damage response. DNA Repair (Amst) 38:94-101

Yoshioka, Ken-ichi; Yoshioka, Yoshiko; Hsieh, Peggy (2006) ATR kinase activation mediated by MutSalpha and MutLalpha in response to cytotoxic O6-methylguanine adducts. Mol Cell 22:501-10

Yang, Yong; Sass, Lauryn E; Du, Chunwei et al. (2005) Determination of protein-DNA binding constants and specificities from statistical analyses of single molecules: MutS-DNA interactions. Nucleic Acids Res 33:4322-34

Schofield, Mark J; Hsieh, Peggy (2003) DNA mismatch repair: molecular mechanisms and biological function. Annu Rev Microbiol 57:579-608

Wang, Hong; Yang, Yong; Schofield, Mark J et al. (2003) DNA bending and unbending by MutS govern mismatch recognition and specificity. Proc Natl Acad Sci U S A 100:14822-7

Selmane, Tassadite; Schofield, Mark J; Nayak, Sunil et al. (2003) Formation of a DNA mismatch repair complex mediated by ATP. J Mol Biol 334:949-65

Schofield, M J; Nayak, S; Scott, T H et al. (2001) Interaction of Escherichia coli MutS and MutL at a DNA mismatch. J Biol Chem 276:28291-9

Junop, M S; Obmolova, G; Rausch, K et al. (2001) Composite active site of an ABC ATPase: MutS uses ATP to verify mismatch recognition and authorize DNA repair. Mol Cell 7:1-12

Schofield, M J; Brownewell, F E; Nayak, S et al. (2001) The Phe-X-Glu DNA binding motif of MutS. The role of hydrogen bonding in mismatch recognition. J Biol Chem 276:45505-8

Biswas, I; Obmolova, G; Takahashi, M et al. (2001) Disruption of the helix-u-turn-helix motif of MutS protein: loss of subunit dimerization, mismatch binding and ATP hydrolysis. J Mol Biol 305:805-16