Crystallization, room-temperature X-ray diffraction and preliminary analysis of Kaposi's sarcoma herpesvirus LANA bound to DNA.
The latency-associated nuclear antigen (LANA) is the latent origin-binding protein and chromatin anchor of the Kaposi's sarcoma herpesvirus (KSHV/HHV-8) genome. Its C-terminal domain (CTD) binds sequence-specifically to the viral origin of replication, whereas the N-terminal domain links it to nucleosomes of cellular chromatin for long-term persistence in dividing host cells. Here, the crystallization and X-ray data acquisition of a mutant LANA CTD in complex with its wild-type target DNA LBS1 is described. This report describes the rational protein engineering for successful co-crystallization with DNA and X-ray diffraction data collection at room temperature on the high-brilliance third-generation synchrotron PETRA III at DESY, Germany.
Department of Structural Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.
Jan
J
Hellert
Department of Structural Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.
Joern
J
Krausze
Institute of Virology, Hannover Medical School, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
Thomas F
TF
Schulz
Department of Structural Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.
Thorsten
T
Lührs
D000956:Antigens, Viral / Q000737:chemistry* / Q000378:metabolism; D003460:Crystallization; D004279:DNA, Viral / Q000737:chemistry* / Q000378:metabolism; D019288:Herpesvirus 8, Human / Q000737:chemistry* / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry* / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012514:Sarcoma, Kaposi / Q000821:virology*; D014961:X-Ray Diffraction
D000956:Antigens, Viral; D004279:DNA, Viral; D009687:Nuclear Proteins; C109540:latency-associated nuclear antigen
Kaposi's sarcoma herpesvirus; latency-associated nuclear antigen; structure-guided mutagenesis