This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The goal of our research is to apply structural biology and high performance computer simulation to investigate molecular mechanisms of bacterial resistance to mercury. Specifically we are interested in solving the first ever X-ray crystal structure of the metal-responsive transcriptional regulator MerR. MerR is the archetype of the MerR family of metalloregulators that controls the transcription of a set of genes (the mer operon) providing Hg resistance in many genera of bacteria isolated from Hg-exposed ecosystems. The mer operon encodes specific genes that facilitate transport of Hg species cleavage of organomercurials and reduction of ionic Hg(II) to volatile elemental Hg(0). The 144-residue MerR binds to its operator DNA and functions as a homodimer. It represses transcription of the mer operon in the absence of Hg(II) and activates transcription upon Hg(II) binding. We wish to obtain a high resolution model to fully characterize Hg(II) binding to MerR. This structural model of MerR will support ongoing neutron scattering and Molecular Dynamics (MD) experiments aimed at studying MerR dynamics and conformational changes upon Hg(II) binding that are essential for understanding its unique mechanism of transcriptional regulation. It is well-known that these multidisciplinary structural studies require maximal resolution X-ray structures of the protein element. As a result a high flux beamline is required to collect high quality data for this proposed research. One day of beamtime (3 shifts) is requested to complete the proposed diffraction studies. We plan to screen crystals of MerR for X-ray diffraction quality and to collect complete redundant data at the highest resolution possible. Structure determination will then be completed by molecular replacement using either a known structure of a paralogous member of the MerR family or by using our current homology model of MerR.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR007707-19
Application #
8363692
Study Section
Special Emphasis Panel (ZRG1-BCMB-P (40))
Project Start
2011-08-01
Project End
2012-07-31
Budget Start
2011-08-01
Budget End
2012-07-31
Support Year
19
Fiscal Year
2011
Total Cost
$6,089
Indirect Cost
Name
University of Chicago
Department
Miscellaneous
Type
Schools of Medicine
DUNS #
005421136
City
Chicago
State
IL
Country
United States
Zip Code
60637
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Cho, Hyun Sun; Schotte, Friedrich; Dashdorj, Naranbaatar et al. (2016) Picosecond Photobiology: Watching a Signaling Protein Function in Real Time via Time-Resolved Small- and Wide-Angle X-ray Scattering. J Am Chem Soc 138:8815-23
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Mariette, Céline; Guérin, Laurent; Rabiller, Philippe et al. (2015) The creation of modulated monoclinic aperiodic composites in n-alkane/urea compounds. Z Kristallogr Cryst Mater 230:5-11
Pfoh, Roland; Pai, Emil F; Saridakis, Vivian (2015) Nicotinamide mononucleotide adenylyltransferase displays alternate binding modes for nicotinamide nucleotides. Acta Crystallogr D Biol Crystallogr 71:2032-9
Sampath, Sujatha; Yarger, Jeffery L (2015) Structural hysteresis in dragline spider silks induced by supercontraction: An x-ray fiber micro-diffraction study. RSC Adv 5:1462-1473
Coppens, Philip; Fournier, Bertrand (2015) New methods in time-resolved Laue pump-probe crystallography at synchrotron sources. J Synchrotron Radiat 22:280-7

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