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PMID: 15738413 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of viral genes essential for replication of murine gamma-herpesvirus 68 using signature-tagged mutagenesis.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 102 ·No. 10 ·2005-03-08 ·Pages 3805-10

Song MJ, Hwang S, Wong WH, Wu TT, Lee S, Liao HI, Sun R

Abstract

Gamma-herpesviruses, Epstein-Barr virus, and Kaposi's sarcoma-associated herpesvirus are important human pathogens, because they are involved in tumor development. Murine gamma-herpesvirus-68 (MHV-68 or gammaHV-68) has emerged as a small animal model system for the study of gamma-herpesvirus pathogenesis and host-virus interactions. To identify the genes required for viral replication in vitro and in vivo, we generated 1,152 mutants using signature-tagged transposon mutagenesis on an infectious bacterial artificial chromosome of MHV-68. Almost every ORF was mutated by random insertion. For each ORF, a mutant with an insertion proximal to the N terminus of each ORF was examined for the ability to grow in fibroblasts. Our results indicate that 41 genes are essential for in vitro growth, whereas 26 are nonessential and 6 attenuated. Replication-competent mutants were pooled to infect mice, which led to the discovery of ORF 54 being important for MHV-68 to replicate in the lung. This genetic analysis of a tumor-associated herpesvirus at the whole genome level validates signature-tagged transposon mutagenesis screening as an effective genetic system to identify important virulent genes in vivo and define interactions with the host immune system.

MeSH Terms
Animals Genes, Viral/physiology Mice Mutagenesis Mutation NIH 3T3 Cells Open Reading Frames Pyrophosphatases/physiology Rhadinovirus/genetics,physiology Virus Replication
Chemicals
Pyrophosphatases dUTP pyrophosphatase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Song Moon Jung
Department of Microbiology, College of Medicine, Hallym University, Chuncheon 200-702, Republic of Korea.
Hwang Seungmin
Wong Wendy H
Wu Ting-Ting
Lee Sangmi
Liao Hsiang-I
Sun Ren
References (44)
44 references, click to expand
  1. Complete sequence and genomic analysis of murine gammaherpesvirus 68.
    J Virol. 1997 Aug;71(8):5894-904 PMID: 9223479
  2. Disruption of the murine gammaherpesvirus 68 M1 open reading frame leads to enhanced reactivation from latency.
    J Virol. 2000 Feb;74(4):1973-84 PMID: 10644370
  3. Four tRNA-like sequences and a serpin homologue encoded by murine gammaherpesvirus 68 are dispensable for lytic replication in vitro and latency in vivo.
    J Gen Virol. 1998 Jan;79 ( Pt 1):149-53 PMID: 9460936
  4. Lung epithelial cells are a major site of murine gammaherpesvirus persistence.
    J Exp Med. 1998 Jun 15;187(12):1941-51 PMID: 9625754
  5. Murine gammaherpesvirus 68: a model for the study of gammaherpesvirus pathogenesis.
    Trends Microbiol. 1998 Jul;6(7):276-82 PMID: 9717216
  6. Macrophages are the major reservoir of latent murine gammaherpesvirus 68 in peritoneal cells.
    J Virol. 1999 Apr;73(4):3273-83 PMID: 10074181
  7. An efficient DNA sequencing strategy based on the bacteriophage mu in vitro DNA transposition reaction.
    Genome Res. 1999 Mar;9(3):308-15 PMID: 10077537
  8. Defined oligonucleotide tag pools and PCR screening in signature-tagged mutagenesis of essential genes from bacteria.
    Biotechniques. 1999 Mar;26(3):473-8, 480 PMID: 10090989
  9. An efficient and accurate integration of mini-Mu transposons in vitro: a general methodology for functional genetic analysis and molecular biology applications.
    Nucleic Acids Res. 1999 Jul 1;27(13):2777-84 PMID: 10373596
  10. Evolution and horizontal transfer of dUTPase-encoding genes in viruses and their hosts.
    J Virol. 1999 Sep;73(9):7710-21 PMID: 10438861
  11. Unraveling immunity to gamma-herpesviruses: a new model for understanding the role of immunity in chronic virus infection.
    Curr Opin Immunol. 1999 Aug;11(4):371-9 PMID: 10448140
  12. The murine gammaherpesvirus 68 v-cyclin is a critical regulator of reactivation from latency.
    J Virol. 2000 Aug;74(16):7451-61 PMID: 10906198
  13. Genomewide function conservation and phylogeny in the Herpesviridae.
    Genome Res. 2001 Jan;11(1):43-54 PMID: 11156614
  14. Natural history of murine gamma-herpesvirus infection.
    Philos Trans R Soc Lond B Biol Sci. 2001 Apr 29;356(1408):569-79 PMID: 11313012
  15. Dissecting the host response to a gamma-herpesvirus.
    Philos Trans R Soc Lond B Biol Sci. 2001 Apr 29;356(1408):581-93 PMID: 11313013
  16. A secreted chemokine binding protein encoded by murine gammaherpesvirus-68 is necessary for the establishment of a normal latent load.
    J Exp Med. 2001 Aug 6;194(3):301-12 PMID: 11489949
  17. Disruption of the M2 gene of murine gammaherpesvirus 68 alters splenic latency following intranasal, but not intraperitoneal, inoculation.
    J Virol. 2002 Feb;76(4):1790-801 PMID: 11799175
  18. Identification of the in vivo role of a viral bcl-2.
    J Exp Med. 2002 Apr 1;195(7):931-40 PMID: 11927636
  19. K3-mediated evasion of CD8(+) T cells aids amplification of a latent gamma-herpesvirus.
    Nat Immunol. 2002 Aug;3(8):733-40 PMID: 12101398
  20. Roles of uracil-DNA glycosylase and dUTPase in virus replication.
    J Gen Virol. 2002 Oct;83(Pt 10):2339-45 PMID: 12237414
  21. The role of retroviral dUTPases in replication and virulence.
    Curr Protein Pept Sci. 2001 Dec;2(4):381-8 PMID: 12374097
  22. Murine gammaherpesvirus 68 lacking thymidine kinase shows severe attenuation of lytic cycle replication in vivo but still establishes latency.
    J Virol. 2003 Feb;77(4):2410-7 PMID: 12551978
  23. Disruption of the gene encoding the gammaHV68 v-GPCR leads to decreased efficiency of reactivation from latency.
    Virology. 2003 Mar 15;307(2):179-90 PMID: 12667789
  24. Disruption of gammaherpesvirus 68 gene 50 demonstrates that Rta is essential for virus replication.
    J Virol. 2003 May;77(10):5731-9 PMID: 12719566
  25. Murid herpesvirus 4 strain 68 M2 protein is a B-cell-associated antigen important for latency but not lymphocytosis.
    J Virol. 2003 Sep;77(17):9700-9 PMID: 12915582
  26. The gammaherpesvirus 68 latency-associated nuclear antigen homolog is critical for the establishment of splenic latency.
    J Virol. 2003 Oct;77(19):10295-303 PMID: 12970414
  27. Functional map of human cytomegalovirus AD169 defined by global mutational analysis.
    Proc Natl Acad Sci U S A. 2003 Oct 14;100(21):12396-401 PMID: 14519856
  28. Transposon-based strategies for microbial functional genomics and proteomics.
    Annu Rev Genet. 2003;37:3-29 PMID: 14616054
  29. ORF73 of murine herpesvirus-68 is critical for the establishment and maintenance of latency.
    J Gen Virol. 2003 Dec;84(Pt 12):3405-16 PMID: 14645921
  30. Identification of proteins associated with murine gammaherpesvirus 68 virions.
    J Virol. 2003 Dec;77(24):13425-32 PMID: 14645600
  31. Functional profiling of a human cytomegalovirus genome.
    Proc Natl Acad Sci U S A. 2003 Nov 25;100(24):14223-8 PMID: 14623981
  32. Murine gammaherpesvirus 68 lacking gp150 shows defective virion release but establishes normal latency in vivo.
    J Virol. 2004 May;78(10):5103-12 PMID: 15113892
  33. Murine gammaherpesvirus 68 open reading frame 31 is required for viral replication.
    J Virol. 2004 Jun;78(12):6610-20 PMID: 15163752
  34. Identification of candidate gammaherpesvirus 68 genes required for virus replication by signature-tagged transposon mutagenesis.
    J Virol. 2004 Oct;78(19):10282-90 PMID: 15367594
  35. Inhibition of phosphorylation of cellular dUTP nucleotidohydrolase as a consequence of herpes simplex virus infection.
    J Cell Biochem. 1990 Aug;43(4):339-53 PMID: 2168898
  36. Herpes simplex virus type 1 dUTPase mutants are attenuated for neurovirulence, neuroinvasiveness, and reactivation from latency.
    J Virol. 1992 Nov;66(11):6706-13 PMID: 1328686
  37. Murine gammaherpesvirus 68 establishes a latent infection in mouse B lymphocytes in vivo.
    J Gen Virol. 1992 Dec;73 ( Pt 12):3275-9 PMID: 1469366
  38. Restoration of cytomegalovirus antigen presentation by gamma interferon combats viral escape.
    J Virol. 1994 Jan;68(1):289-97 PMID: 8254740
  39. Simultaneous identification of bacterial virulence genes by negative selection.
    Science. 1995 Jul 21;269(5222):400-3 PMID: 7618105
  40. Mature B cells are required for acute splenic infection, but not for establishment of latency, by murine gammaherpesvirus 68.
    J Virol. 1996 Oct;70(10):6775-80 PMID: 8794315
  41. Identification of Staphylococcus aureus virulence genes in a murine model of bacteraemia using signature-tagged mutagenesis.
    Mol Microbiol. 1997 Oct;26(2):399-407 PMID: 9383163
  42. Host and viral genetics of chronic infection: a mouse model of gamma-herpesvirus pathogenesis.
    Curr Opin Microbiol. 1999 Aug;2(4):403-9 PMID: 10458986
  43. In vivo genetic analysis of bacterial virulence.
    Annu Rev Microbiol. 1999;53:129-54 PMID: 10547688
  44. Genetic content and preliminary transcriptional analysis of a representative region of murine gammaherpesvirus 68.
    J Gen Virol. 1997 Jun;78 ( Pt 6):1425-33 PMID: 9191940
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0404521102
Published
2005-03-08
Epub
2005-00-28
Pages
3805-10
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC553290
Subset
IM
Grants
NCI NIH HHS · CA 83525 · United States
NCI NIH HHS · CA 91791 · United States
NIDCR NIH HHS · R01 DE014153 · United States
NIDCR NIH HHS · DE 14153 · United States
NCI NIH HHS · R01 CA091791 · United States
External Links
PubMed source DOI: 10.1073/pnas.0404521102 PMC Full Text View journal details
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