Volume 8 Supplement 1

15th International Conference on Human Retroviruses: HTLV and Related Viruses

Open Access

In vitro assembly of xenotropic murine leukemia virus-related virus CA-NC protein

  • Romana Hadravová1,
  • Jitka Štokrová1,
  • Michal Doležal1,
  • Iva Pichová1,
  • Tomáš Ruml2 and
  • Michaela Rumlová1Email author
Retrovirology20118(Suppl 1):A236

DOI: 10.1186/1742-4690-8-S1-A236

Published: 6 June 2011

Using in vitro expression/assembly system we studied the formation of virus-like particles Xenotropic Murine Leukemia Virus-related virus (XMRV). XMRV is novel human gammaretrovirus discovered in association with human prostate tumors. The genome organization is typical for gammaretroviruses consisting of two overlapping ORFs coding for Gag-Pro-Pol and Env polyproteins. The predicted Gag polyprotein consists of 536 amino acids and is separated from the Pro-Pol sequence by UAG stop codon.

Based on the amino acids similarities between MLV and XMRV Gag polyproteins, we designed primers bordering CA-NC region of Gag. Resulting PCR fragment was cloned into pET22b vector for expression of CA-NC in E. coli. We found that purified XMRV full-length CANC, starting with the conserved proline residue at the N-terminus of CA, was not able to assemble into particles. However, a modification of the N-terminus of CANC (modCANC) enabled formation of spherical particles. Moreover, the negative staining of the in vitro assembled particles of XMRV modCANC revealed different organization of protein layers in comparison to CA-NC of M-PMV.

Authors’ Affiliations

(1)
Department of Biochemistry, Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences
(2)
Department of Biochemistry and Microbiology, Institute of Chemical Technology

Copyright

© Hadravová et al; licensee BioMed Central Ltd. 2011

This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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