Elsevier

Journal of Structural Biology

Volume 108, Issue 3, May–June 1992, Pages 209-220
Journal of Structural Biology

Distribution of viral RNA molecules during the adenovirus type 5 infectious cycle in HeLa cells

https://doi.org/10.1016/1047-8477(92)90021-2Get rights and content

Abstract

Viral RNA was localized ultrastructurally by in situ hybridization with a biotinylated viral DNA probe and colloidal gold label in HeLa cells during infection with adenovirus type 5. Transcription was monitored by high-resolution autoradiography after short pulses with tritiated uridine. At the earliest stage of virus-induced nuclear transformation, viral RNA was restricted to the small compact fibrillar “early replicative sites” which we had previously demonstrated to be the site of viral DNA replication (Puvion-Dutilleul and Puvion, 1990b). Protease-DNase-treated sections revealed that these fibrillar masses rapidly enlarged and gave rise to a juxtaposed loose fibrillogranular structure devoid of viral RNA. Subsequently, the function of the compact fibrillar zones changed to become the sites of accumulation of single-stranded (ss) viral DNA molecules, and the contiguous new fibrillogranular zones (previously named the peripheral replicative zones, Puvion-Dutilleul and Puvion, 1990a) became not only the centers of replicating viral double-stranded DNA but also the only sites of viral RNA molecules including the elongating RNA transcripts. Pulse-chase experiments revealed an unexpected accumulation of RNA molecules in these extranucleolar regions of infected nuclei.

References (22)

  • A.L. Beyer et al.

    J. Mol. Biol

    (1981)
  • N. Ledinko

    Virology

    (1972)
  • J.B. Lewis et al.

    Cell

    (1977)
  • J.R. Nevins et al.

    Cell

    (1978)
  • F. Puvion-Dutilleul et al.

    J. Struct. Biol

    (1990)
  • F. Puvion-Dutilleul et al.

    Biol. Cell

    (1991)
  • H.J. Raskas et al.

    Virology

    (1970)
  • D.J. Wolgemuth et al.

    J. Mol. Biol

    (1981)
  • E.B. Ziff
  • M. Bendayan

    J. Electron Microsc. Tech

    (1984)
  • M. Bouteille

    J. Microsc. Biol. Cell

    (1976)
  • Cited by (43)

    • The role of Cajal bodies in the expression of late phase adenovirus proteins

      2010, Virology
      Citation Excerpt :

      Transcription sites overlap the replication foci but extend further into the nucleoplasm away from E2A-72K domains, forming patterns of inter-connecting rings (Pombo et al., 1994). The transcription zones also contain nascent viral RNA (Pombo et al., 1994; Puvion-Dutilleul and Puvion, 1992), splicing snRNPs (Aspegren and Bridge, 2002), viral introns and exons (Rebelo et al., 1996; Aspegren et al., 1998; Bridge et al., 1996). The transition from the early to the late phase requires viral gene expression from replicated templates (Chalberg and Kelley, 1989).

    • An early function of the adenoviral E1B 55 kDa protein is required for the nuclear relocalization of the cellular p53 protein in adenovirus-infected normal human cells

      2008, Virology
      Citation Excerpt :

      Moreover, the E1B 55 kDa and E4 Orf6 proteins form a complex that can associate with cellular proteins to form an infected cell-specific, cullin5-containing E3 ubiquitin ligase that polyubiquitinylates and stimulates proteasomal degradation of p53 (Harada et al., 2002; Querido et al., 2001; Sarnow et al., 1984; Woo and Berk, 2007). In infected cell nuclei, the E1B 55 kDa and E4 Orf6 proteins colocalize to the peripheral zones of specific nuclear microenvironments where viral DNA replication, transcription of viral late genes and the initial posttranscriptional processing of viral late mRNAs take place (Aspegren et al., 1998; Bridge et al., 1995; Ornelles and Shenk, 1991; Pombo et al., 1994; Puvion-Dutilleul et al., 1992). Mutations that reduce binding of the E1B 55 kDa (E1B) to the E4 Orf6 protein result in altered localization of E1B, and inefficient export of viral late mRNAs (Gonzalez and Flint, 2002; Ornelles and Shenk, 1991; Rubenwolf et al., 1997), correlating efficient viral late mRNA export with the organization of infected cell nuclei.

    View all citing articles on Scopus

    This work was supported by general grants from the Centre National de la Recherche Scientifique and special grants from the Association pour la Recherche sur le Cancer (Villejuif/France) and the Fondation pour la Recherche Médicale.

    3

    F. Puvion-Dutilleul is a member of the Institut National de la Santé et de la Recherche médicale.

    2

    Present address: Institute of General and Comparative Pathology, Bulgarian Academy of Sciences, Acad. G. Bonchev, Str. Bl. 25, Sofia 1113, Bulgaria.

    View full text