1887

Journal of General Virology header logo

Volume 74, Issue 7

Intranuclear foci containing low abundance herpes simplex virus latency-associated transcripts visualized by non-isotopic hybridization Free

Abstract

During latent infection of neurons with herpes simplex virus type 1 (HSV-1), several RNA transcripts of varying abundance arise from a single locus within the virus repeats. The functions of latency-associated transcripts (LATs) are unknown and the relationship between the various RNA species requires further clarification. Reported here is a novel approach to the study of HSV transcripts during latency, based on the increasing realization that cellular and viral RNAs are synthesized and processed by macromolecular complexes that occupy discrete compartments within the nucleoplasm of a cell. High resolution non-isotopic hybridization was used to study the intranuclear topology of HSV-1 LATs in primary sensory neurons of latently infected mice and humans. Low abundance (minor) LATs were localized to sharply defined intranuclear foci of 1 to 3 μm in diameter. On average, there were 2·6 to 2·8 foci/LAT neuronal profile (5 μm), representing 13 to 14 foci/cell. In contrast to the focal deployment of minor LATs, the more abundant latency-associated RNAs were distributed diffusely throughout the nucleoplasms of latency infected neurons, with prominent sparing of nucleolar regions. These data establish a foundation for studying the synthesis, processing and transport of LATs . It should now be possible to investigate the nature of those cellular products which associate with HSV-1 encoded LATs and thereby determine whether minor LATs are associated with previously characterized macromolecular complexes, such as those responsible for processing of pre-messenger RNA.

  • Received:
  • Accepted:
  • Published Online:
© Journal of General Virology 1993
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-74-7-1363
1993-07-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/74/7/JV0740071363.html?itemId=/content/journal/jgv/10.1099/0022-1317-74-7-1363&mimeType=html&fmt=ahah

References

  1. Carmo-Fonseca M, Pepperkok R, Sproat BS, Ansorge W, Swanson MS, Lamond AI. 1991a; In vivo detection of snRNP-rich organelles in the nuclei of mammalian cells.. EMBO Journal 10:1863–1873
     [Google Scholar]
  2. Carmo-Fonseca M, Tollervey D, Pepperkok R, Barabino SML, Merdes A, Brunner C, Zamore PD, Green MR, Hurt E, Lamond AI. 1991b; Mammalian nuclei contain foci which are highly enriched in components of the pre-mRNA splicing machinery.. EMBO Journal 10:195–206
     [Google Scholar]
  3. Carter KC, Lawrence JB. 1991; DNA and RNA within the nucleus: how much spatial organization?. Journal of Cellular Biochemistry 47:124–129
     [Google Scholar]
  4. Carter KC, Taneja KL, Lawrence JB. 1991; Discrete nuclear domains of poly(A) RNA and their relationship to the functional organization of the nucleus.. Journal of Cell Biology 115:1191–1202
     [Google Scholar]
  5. Devi-Rao G, Goodart SA, Hecht LM, Rochford R, Rice MK, Wagner EK. 1991; Relationship between polyadeny-lated and nonpolyadenylated herpes simplex virus type 1 latency-associated transcripts.. Journal of Virology 65:2179–2190
     [Google Scholar]
  6. Dobson AT, Sederati F, Devi-Rao G, Flanagan WM, Farrell MJ, Stevens JG, Wagner EK, Feldman LT. 1989; Identification of the latency-associated transcript promoter by expression of rabbit beta-globin mRNA in mouse sensory nerve ganglia latently infected with a recombinant herpes simplex virus.. Journal of Virology 63:3844–3851
     [Google Scholar]
  7. Farrell MJ, Dobson AT, Feldman LT. 1991; Herpes simplex virus latency-associated transcript is a stable intron.. Proceedings of the National Academy of Sciences, U.S.A. 88:790–794
     [Google Scholar]
  8. Gowans EJ, Jilbert AR, Burrell CJ. 1989; Detection of specific DNA and RNA sequences in tissues and cells by in situ hybridization.. Nucleic Acid Probe139–158Edited by Symons RH. Boca Raton: CRC Press.; In pp
    [Google Scholar]
  9. Hill TJ, Field HJ, Blyth WA. 1975; Acute and recurrent infection with herpes simplex virus in the mouse: a model for studying latency and recurrent disease.. Journal of General Virology 28:341–353
     [Google Scholar]
  10. Ho DY. 1992; Herpesvirus latency : molecular aspects.. Progress in Medical Virology 39:76–116
     [Google Scholar]
  11. Lawrence JB, Singer RH, Marselle LM. 1989; Highly localized tracks of specific transcripts within interphase nuclei visualized by in situ hybridization.. Cell 57:493–502
     [Google Scholar]
  12. Lawrence JB, Marselle LM, Byron KS, Johnson CV, Sullivan JL, Singer RH. 1990; Subcellular localization of low-abundance human immunodeficiency virus nucleic acid sequences visualized by fluorescence in situ hybridization.. Proceedings of the National Academy of Sciences, U.S.A. 249:928–932
     [Google Scholar]
  13. McLean IW, Nakane PK. 1974; Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy.. Journal of Histochemistry and Cytochemistry 22:1077–1083
     [Google Scholar]
  14. Mitchell WJ, Lirette RP, Fraser NW. 1990a; Mapping of low abundance latency-associated mRNA in the trigeminal ganglia of mice latently infected with herpes simplex virus type 1.. Journal of General Virology 7:125–132
     [Google Scholar]
  15. Mitchell WJ, Steiner I, Brown SM, MacLean AR, Subak-Sharpe JH, Fraser NW. 1990b; A herpes simplex virus type 1 variant, deleted in the promoter region of the latency-associated transcripts, does not produce any detectable minor RNA species during latency in the mouse trigeminal ganglion.. Journal of General Virology 71:953–957
     [Google Scholar]
  16. Nevins JR. 1983; The pathway of eukaryotic mRNA formation.. Annual Reviews of Biochemistry 52:441–466
     [Google Scholar]
  17. Rødahl E, Stevens JG. 1992; Differential accumulation of herpes simplex type 1 latency-associated transcripts in sensory autonomic ganglia.. Virology 189:385–388
     [Google Scholar]
  18. Russell WC. 1962; A sensitive and precise assay for herpes virus.. Nature, London 195:1028–1029
     [Google Scholar]
  19. Simmons A, La Vista AB. 1989; Neural infection in mice after cutaneous inoculation with HSV-1 is under complex host genetic control.. Virus Research 13:263–270
     [Google Scholar]
  20. Simmons A, Nash AA. 1984; Zosteriform spread of herpes simplex virus as a model of recrudescence and its use to investigate the role of immune cells in prevention of recurrent disease.. Journal of Virology 52:816–821
     [Google Scholar]
  21. Simmons A, Slobedman B, Speck P, Arthur J, Efstathiou S. 1992; Two patterns of persistence of herpes simplex virus DNA sequences in the nervous systems of latently infected mice.. Journal of General Virology 73:1287–1291
     [Google Scholar]
  22. Speck P, Simmons A. 1991; Divergent molecular pathways of productive and latent infection with a virulent strain of herpes simplex virus type.. Journal of Virology 65:4001–4005
     [Google Scholar]
  23. Spivack JG, Fraser NW. 1987; Detection of herpes simplex virus type 1 transcripts during latent infection in mice.. Journal of Virology 61:3841–3847
     [Google Scholar]
  24. Spivack JG, Woods GM, Fraser NW. 1991; Identification of a novel latency-specific splice donor signal within the herpes simplex virus type 1 2 0 kilobase latency-associated transcript (LAT): translation inhibition of LAT open reading frames by the intron within the 2 0-kilobase LAT.. Journal of Virology 65:6800–6810
     [Google Scholar]
  25. Stevens JG. 1989; Human herpesviruses: a consideration of the latent state.. Microbiological Reviews 53:318–332
     [Google Scholar]
  26. Stevens JG, Wagner EK, Devi-Rao GB, Cook ML, Feldman LT. 1987; RNA complementary to a herpesvirus a gene mRNA is prominent in latently infected neurons.. Science 235:1056–1059
     [Google Scholar]
  27. Wagner EK, Devi-Rao G, Feldman LT, Dobson AT, Zhang YF, Flanagan WM, Stevens JG. 1988a; Physical characterization of the herpes simplex virus latency-associated transcript in neurons.. Journal of Virology 62:1194–1202
     [Google Scholar]
  28. Wagner EK, Flanagan WM, Devi-Rao GB, Zhang YF, Hill JM, Anderson KP, Stevens JG. 1988b; The herpes simplex virus latency-associated transcript is spliced during the latent phase of infection.. Journal of Virology 62:4577–4585
     [Google Scholar]
  29. Wang J, Cao LG, Wang YL, Pederson T. 1991; Localization of pre-messenger RNA at discrete nuclear sites.. Proceedings of the National Academy of Sciences, U.S.A. 88:7391–7395
     [Google Scholar]
  30. Wechsler SL, Nesburn AB, Watson R, Slanina S, Ghiasi H. 1988; Fine mapping of the major latency-related gene of herpes simplex virus type 1: alternative splicing produces distinct latency-related RNAs containing open reading frames.. Journal of Virology 88:4051–4058
     [Google Scholar]
  31. Wildy P, Field HJ, Nash AA. 1982; Classical herpes latency revisited.. Virus Persistenc133–167Edited by Mahy BWJ, Minson AC, Darby GK. Cambridge: Cambridge University Press.;
     [Google Scholar]
  32. Xing Y, Lawrence J. B. 1991; Preservation of specific RNA distribution within the chromatin-depleted nuclear substructure demonstrated by in situ hybridization coupled with biochemical fractionation.. Journal of Cell Biology 112:1055–1063
     [Google Scholar]
  33. Zwaagstra J, Ghiasi H, Nesburn AB, Wechsler SL. 1989; In vitro promoter activity associated with the latency-associated transcript gene of herpes simplex virus type 1.. Journal of General Virology 70:2163–2169
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-74-7-1363
Loading
Intranuclear foci containing low abundance herpes simplex virus latency-associated transcripts visualized by non-isotopic in situ hybridization
J. Gen. Virol. 74, 1363 (1993); https://doi.org/10.1099/0022-1317-74-7-1363
/content/journal/jgv/10.1099/0022-1317-74-7-1363
/content/journal/jgv/10.1099/0022-1317-74-7-1363
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error