Replication protein A (RPA) is a key component of the DNA replication and repair machineries. RPA is a single stranded DNA binding protein that interacts with DNA replication orgin binding proteins, DNA polymerases, transcription activator proteins and DNA repair factors, including those that are mutated in several forms of human cancer. RPA is a heterotrimer; the DNA binding domain has been localized to the largest subunit. We have used limited proteolysis and deletion analysis to identify the minimal single strand DNA binding domain. We have cryslalized this domain together with ssDNA. Cryo-protected crystals diffract to a resolution of 2.2A on our RAXIS-II equipped with double double focussing mirrors. We are pursuing heavy atom searches on our unit and have prepared selenomethionine-labelled protein for co-crystallization. In this proposal, we wish to use, collect higher resolution native as well as heavy atom data sets.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001646-20
Application #
6667789
Study Section
Project Start
2002-09-30
Project End
2003-08-14
Budget Start
Budget End
Support Year
20
Fiscal Year
2002
Total Cost
$142,703
Indirect Cost
Name
Cornell University
Department
Type
DUNS #
City
Ithaca
State
NY
Country
United States
Zip Code
14850
Kozlov, Guennadi; Wong, Kathy; Gehring, Kalle (2018) Crystal structure of the Legionella effector Lem22. Proteins 86:263-267
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Xu, Jie; Kozlov, Guennadi; McPherson, Peter S et al. (2018) A PH-like domain of the Rab12 guanine nucleotide exchange factor DENND3 binds actin and is required for autophagy. J Biol Chem 293:4566-4574
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Chen, Yu Seby; Kozlov, Guennadi; Fakih, Rayan et al. (2018) The cyclic nucleotide-binding homology domain of the integral membrane protein CNNM mediates dimerization and is required for Mg2+ efflux activity. J Biol Chem 293:19998-20007
Xu, Caishuang; Kozlov, Guennadi; Wong, Kathy et al. (2016) Crystal Structure of the Salmonella Typhimurium Effector GtgE. PLoS One 11:e0166643
Cogliati, Massimo; Zani, Alberto; Rickerts, Volker et al. (2016) Multilocus sequence typing analysis reveals that Cryptococcus neoformans var. neoformans is a recombinant population. Fungal Genet Biol 87:22-9
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Lucido, Michael J; Orlando, Benjamin J; Vecchio, Alex J et al. (2016) Crystal Structure of Aspirin-Acetylated Human Cyclooxygenase-2: Insight into the Formation of Products with Reversed Stereochemistry. Biochemistry 55:1226-38
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