A Role for Transportin in the Nuclear Import of Adenovirus Core Proteins and DNA

Hindley, Clemence E.; Lawrence, Fiona; Matthews, David A.

doi:10.1111/j.1600-0854.2007.00618.x

Cited by 58 publications

(76 citation statements)

References 24 publications

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“…6), as reported for wild-type pV (29,32). The absence of pV or GFP-pV on the viral cores in the nucleus is not surprising, since earlier studies showed that the viral DNA and pVII alone can be a template for replication and transcription without pV (9,28,67).…”

Section: Vol 85 2011mentioning

confidence: 69%

Stepwise Loss of Fluorescent Core Protein V from Human Adenovirus during Entry into Cells

Püntener

Engelke

Ruzsics

et al. 2011

J Virol

112

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Human adenoviruses (Ads) replicate and assemble particles in the nucleus. They organize a linear double-strand DNA genome into a condensed core with about 180 nucleosomes, by the viral proteins VII (pVII), pX, and pV attaching the DNA to the capsid. Using reverse genetics, we generated a novel, nonconditionally replicating Ad reporter by inserting green fluorescent protein (GFP) at the amino terminus of pV. Purified Ad2-GFP-pV virions had an oversized complete genome and incorporated about 38 GFP-pV molecules per virion, which is about 25% of the pV levels in Ad2. GFP-pV cofractionated with the DNA core, like pV, and newly synthesized GFP-pV had a subcellular localization indistinguishable from that of pV, indicating that GFP-pV is a valid reporter for pV. Ad2-GFP-pV completed the replication cycle, although at lower yields than Ad2. Incoming GFP-pV (or pV) was not imported into the nucleus. Virions lost GFP-pV at two points during the infection process: at entry into the cytosol and at the nuclear pore complex, where capsids disassemble. Disassembled capsids, positive for the conformation-specific antihexon antibody R70, were devoid of GFP-pV. The loss of GFP-pV was reduced by the macrolide antibiotic leptomycin B (LMB), which blocks nuclear export and adenovirus attachment to the nuclear pore complex. LMB inhibited the appearance of R70 epitopes on Ad2 and Ad2-GFP-pV, indicating that the loss of GFP-pV from Ad2-GFP-pV is an authentic step in the adenovirus uncoating program. Ad2-GFP-pV is genetically complete and hence enables detailed analyses of infection and spreading dynamics in cells and model organisms or assessment of oncolytic adenoviral potential.DNA viruses and retroviruses maintain and replicate their genomes in host cell nuclei by using histone-based nucleosomes, similar to chromatin, or they encode their own DNA binding and DNA-organizing proteins (34,45,47). They assemble and maintain their genomes in different chromatin states by packaging the nucleic acids into proteinaceous capsids and sometimes lipid envelopes and thereby traffic their genome within and transmit it between cells (8, 41). The simian virus 40 (SV40) polyomavirus, for example, packages its virion DNA with cellular core histones and uses histones to replicate in infected nuclei (19). Herpesviruses, on the other hand, condense their double-strand DNA in particles with the help of polyamines and use histones during latent residence within infected nuclei or use irregularly spaced nucleosomes during productive phases of infection (45).Adenoviruses (Ads) replicate and assemble particles in the nucleus. They encode their own histone-like proteins to condense a linear double-strand DNA genome of about 36 kbp into a proteinaceous DNA core. Although it is unknown how the viral DNA is precisely organized in the virion, isolated cores of species C human adenovirus serotypes 2 and 5 (Ad2/5) contain six viral proteins, the basic proteins V (pV), pVII, and pX; the terminal protein covalently attached to the 5Ј ends of the DNA; and small numbe...

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Section: Vol 85 2011mentioning

confidence: 69%

Stepwise Loss of Fluorescent Core Protein V from Human Adenovirus during Entry into Cells

Püntener

Engelke

Ruzsics

et al. 2011

J Virol

112

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“…Since it has been shown by yeast two-hybrid analysis that protein V has an affinity with a cellular protein, p32, which shuttles between the nucleus and mitochondria, it may be that there are a number of routes which the invading virus template can hijack to reach the nucleus (Matthews & Russell, 1998a). A recent paper suggests that protein VII is the major mediator of virus DNA import into the nucleus, facilitated primarily by transportin (Hindley et al, 2007). There is also evidence that, prior to nuclear import, V and VII interact with the Golgi apparatus via the MTOC and then travel to the endoplasmic reticulum prior to nuclear import (C. Hindley and D. A. Matthews, personal communication).…”

Section: Core Proteins V Vii Mu Protease and Tpmentioning

confidence: 99%

Adenoviruses: update on structure and function

Russell

2009

Journal of General Virology

344

268

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“…The nuclear import of recombinant mature protein VII is mediated by the NTF transportin-1 [27], which also mediates the nuclear import of adenoviral DNA [27] and exogenous DNA [28]. However, in addition to transportin-1, the NTFs importin-b and importin-7 also play a role in adenovirus DNA nuclear import [14,27,29].…”

Section: Introductionmentioning

confidence: 99%