Crystallization, room-temperature X-ray diffraction and preliminary analysis of Kaposi's sarcoma herpesvirus LANA bound to DNA

Acta Crystallogr F Struct Biol Commun. 2014 Nov;70(Pt 11):1570-4. doi: 10.1107/S2053230X14019906. Epub 2014 Oct 25.

Abstract

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.

Keywords: Kaposi's sarcoma herpesvirus; latency-associated nuclear antigen; structure-guided mutagenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Viral / chemistry*
  • Antigens, Viral / metabolism
  • Crystallization
  • DNA, Viral / chemistry*
  • DNA, Viral / metabolism
  • Herpesvirus 8, Human / chemistry*
  • Herpesvirus 8, Human / metabolism
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / metabolism
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Sarcoma, Kaposi / virology*
  • X-Ray Diffraction

Substances

  • Antigens, Viral
  • DNA, Viral
  • Nuclear Proteins
  • latency-associated nuclear antigen