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Structure and dynamics in proteins: Part I. structural origins of specific DNA recognition by GFI-1 Part II. structural and dynamic studies of γS-crystallin and OPJ, implications for cataract formation

Lee, Soojin
http://rave.ohiolink.edu/etdc/view?acc_num=osu1189025356

Abstract Details

2007, Doctor of Philosophy, Ohio State University, Biophysics.
Gfi-1 is a transcriptional repressor crucial for precise regulation of cell proliferation and differentiation in hematopoiesis. Recently, it has also been demonstrated to be capable of restricting the proliferation of hematopoietic stem cells (HSCs), a process which appears to be vital for the long term competency of HSCs. These two seemingly opposite outcomes of regulation are likely to arise from interactions with a variety of different cellular partners. Such protein-protein interactions can directly affect the genes Gfi-1 recognizes through its DNA binding zinc-finger domain. Interestingly, biochemical foot-printing studies by methylation suggested a novel minor groove DNA-binding model by these canonical C2H2 zinc fingers. The focus of the work herein is to determine the solution structure of Gfi-1 zinc fingers 3-5 (Gfizf35) bound to a 16-mer consensus DNA to provide insights into the mechanism of specific DNA recognition by Gfi-1. The structure of Gfizf35 bound with its consensus DNA has been solved by using multidimensional NMR, including NOE-based distance restraints, torsion angle restraints, and residual dipolar couplings from both the protein and the DNA. Unlike the proposed minor groove model, the solution structure reveals that Gfi-1 zinc fingers wrap around the target DNA by binding the major groove. Protein-DNA interactions are mediated by residues located on the entire outer-surface of the α-helices, the second β-strands and the linkers between them. The last zinc finger recognizes the AATC core sequence of the consensus site by forming specific hydrogen bonds between the side chains of Asn72, Gln69 and the two invariant adenosine bases. Similarly, the fourth zinc finger also exhibits specific interaction between Gln41 and a less conserved but still important adenosine base. The structure furnishes insights into the mechanism of specific DNA recognition by Gfi-1 and provides a rationale for understanding the disease-caused by N72S mutation, mutagenesis and methylation data.
Zhengrong (Justin) Wu (Advisor)
Mark Foster (Other)
Venkat Gopalan (Other)
Dongping Zhong (Other)
Biophysics, General

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Citations

  • Lee, S. (2007). Structure and dynamics in proteins: Part I. structural origins of specific DNA recognition by GFI-1 Part II. structural and dynamic studies of γS-crystallin and OPJ, implications for cataract formation [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1189025356

    APA Style (7th edition)

  • Lee, Soojin. Structure and dynamics in proteins: Part I. structural origins of specific DNA recognition by GFI-1 Part II. structural and dynamic studies of γS-crystallin and OPJ, implications for cataract formation. 2007. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1189025356.

    MLA Style (8th edition)

  • Lee, Soojin. "Structure and dynamics in proteins: Part I. structural origins of specific DNA recognition by GFI-1 Part II. structural and dynamic studies of γS-crystallin and OPJ, implications for cataract formation." Doctoral dissertation, Ohio State University, 2007. http://rave.ohiolink.edu/etdc/view?acc_num=osu1189025356

    Chicago Manual of Style (17th edition)

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© 2007, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.