BIOCHEMICAL AND CRYSTALLOGRAPHIC STUDIES OF DNA REPLICATION AND PACKAGING PROTEIN NS1 FROM HUMAN PARVOVIRUSES

picture of Daniel Koblas presenting his/her poster: BIOCHEMICAL AND CRYSTALLOGRAPHIC STUDIES OF DNA REPLICATION AND PACKAGING PROTEIN NS1 FROM HUMAN PARVOVIRUSES

Daniel Koblas , Sunil Tewary, Liang Tang, Department of Molecular Biosciences, University of Kansas, Lawrence KS

BIOCHEMICAL AND CRYSTALLOGRAPHIC STUDIES OF DNA REPLICATION AND PACKAGING PROTEIN NS1 FROM HUMAN PARVOVIRUSES

The parvovirus Human Bocavirus (HBoV) was identified in 2005 and is associated with acute respiratory illness and pneumonia worldwide.  Human parvoviruses such as HBoV form infectious particles consisting of a non-enveloped icosahedral protein capsid approximately 25 nanometers in diameter that is packaged with the single-stranded viral DNA.  During virus assembly, a protein complex forms a molecular motor that packages the DNA into the pre-formed capsid through a pore present at the 5-fold vertices of the capsid.  Non-Structural Protein (NS1) is a multifunctional protein from certain genera of human parvoviruses that plays a critical role in viral single-stranded DNA replication and is also responsible for packaging the viral DNA into the pre-formed capsid during virus assembly.  The DNA binding/nickase activities and helicase/ATPase activities of the N-terminal and central domains, respectively, of the NS1 protein are essential in these processes, though the precise mechanisms by which these activities are carried out are not well known.  In this study, NS1 N-terminal domain from Human Bocavirus was overexpressed in E. coli, purified to high homogeneity with Ni-NTA chromatography and gel filtration chromatography, and crystallized using MPD as the precipitant.  X-ray diffraction data were collected using synchrotoron radiation and processed at a 2.6 Å resolution.  The crystal belonged to the tetragonal space group P422 with unit cell parameters a=200 and c=56 Å.

The research in Dr. Tang's laboratory has been supported by the grant R01GM090010 from the National Institute of General Medical Sciences of the National Institutes of Health. Daniel Koblas is supported by the National Science Foundation Research Experience for Undergraduates summer program.

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