During the last year we have solved two new virus structures and we are on the verge of solving a third with data collected at CHESS. Three virus structures related to the RNA insect pathogen Flock House Virus (FHV) were also determined with CHESS data. We have generated a 5? electron density map of the first dsDNA bacteriophage capsid, HK97 (P21 a=580, b=625, c=790?, beta=90.0o). This structure, currently being extended to 3.5? resolution, is the first example of a particle that undergoes a large scale maturation polymorphism. Our structure is of the mature head which is ~650? in diameter. This map was the result of collecting two different data sets (100? resolution to 7? and 20? resolution to 4.3?) at CHESS with exceptional help and support from MacCHESS personnel. We also solved the structure of cricket paralysis virus (CrPV) which is the first example of a picorna-like insect virus. This structure is being refined at 2.4? resolution. Unlike normal mamallian picornaviruses, the structural proteins are at the 3 end of the RNA and the VP4 chain is cleaved from the N terminus of VP3, not from the N terminus of VP2. Finally we are on the verge of solving the structure of rice yellow mottle virus (RYMV), the cause of one of the most devasting losses in food crops in Africa. Finally, we solved four mutants of FHV that are virus-like particles produced in a baculovirus expression system. The first was the wild type recombinant particle, the second a non cleaving mutant, the third an assembly mutant that produced a statistical distribution of particles including a T=3 particle that crsyatllized and diffracted x-rays to 2.4?. All three structures have lead to new understanding of the viral biology.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR001646-20
Application #
6667803
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
Ménade, Marie; Kozlov, Guennadi; Trempe, Jean-François et al. (2018) Structures of ubiquitin-like (Ubl) and Hsp90-like domains of sacsin provide insight into pathological mutations. J Biol Chem 293:12832-12842
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
Dean, Dexter N; Rana, Pratip; Campbell, Ryan P et al. (2018) Propagation of an A? Dodecamer Strain Involves a Three-Step Mechanism and a Key Intermediate. Biophys J 114:539-549
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
Oot, Rebecca A; Kane, Patricia M; Berry, Edward A et al. (2016) Crystal structure of yeast V1-ATPase in the autoinhibited state. EMBO J 35:1694-706
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
Bauman, Joseph D; Harrison, Jerry Joe E K; Arnold, Eddy (2016) Rapid experimental SAD phasing and hot-spot identification with halogenated fragments. IUCrJ 3:51-60

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