Nuclear Targeting of Adenovirus Type 2 Requires CRM1-mediated Nuclear Export

Incoming adenovirus type 2 (Ad2) and Ad5 shuttle bidirectionally along microtubules, biased to the microtubule-organizing center by the dynein/dynactin motor complex. It is unknown how the particles reach the nuclear pore complex, where capsids disassemble and viral DNA enters the nucleus. Here, we identified a novel link between nuclear export and microtubule-mediated transport. Two distinct inhibitors of the nuclear export factor CRM1, leptomycin B (LMB) and ratjadone A (RJA) or CRM1-siRNAs blocked adenovirus infection, arrested cytoplasmic transport of viral particles at the microtubule-organizing center or in the cytoplasm and prevented capsid disassembly and nuclear import of the viral genome. In mitotic cells where CRM1 is in the cytoplasm, adenovirus particles were not associated with microtubules but upon LMB treatment, they enriched at the spindle poles implying that CRM1 inhibited microtubule association of adenovirus. We propose that CRM1, a nuclear factor exported by CRM1 or a protein complex containing CRM1 is part of a sensor mechanism triggering the unloading of the incoming adenovirus particles from microtubules proximal to the nucleus of interphase cells.

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Faculty Opinions recommendation of Nuclear targeting of adenovirus type 2 requires CRM1-mediated nuclear export.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1026285.325010 ◽

2005 ◽

Author(s):

Greg Smith

Keyword(s):

Nuclear Export ◽

Adenovirus Type ◽

Nuclear Targeting ◽

Adenovirus Type 2

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Cytoplasmic transport and nuclear import of plasmid DNA

Bioscience Reports ◽

10.1042/bsr20160616 ◽

2017 ◽

Vol 37 (6) ◽

Cited By ~ 34

Author(s):

Haiqing Bai ◽

Gillian M. Schiralli Lester ◽

Laura C. Petishnok ◽

David A. Dean

Keyword(s):

Nuclear Envelope ◽

Model Systems ◽

Nuclear Pore ◽

Nuclear Pore Complexes ◽

Microtubule Organizing Center ◽

Microtubule Network ◽

Naked Dna ◽

Cytoplasmic Transport ◽

Organizing Center ◽

Pore Complexes

Productive transfection and gene transfer require not simply the entry of DNA into cells and subsequent transcription from an appropriate promoter, but also a number of intracellular events that allow the DNA to move from the extracellular surface of the cell into and through the cytoplasm, and ultimately across the nuclear envelope and into the nucleus before any transcription can initiate. Immediately upon entry into the cytoplasm, naked DNA, either delivered through physical techniques or after disassembly of DNA–carrier complexes, associates with a large number of cellular proteins that mediate subsequent interactions with the microtubule network for movement toward the microtubule organizing center and the nuclear envelope. Plasmids then enter the nucleus either upon the mitotic disassembly of the nuclear envelope or through nuclear pore complexes in the absence of cell division, using a different set of proteins. This review will discuss our current understanding of these pathways used by naked DNA during the transfection process. While much has been elucidated on these processes, much remains to be discerned, but with the development of a number of model systems and approaches, great progress is being made.

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Morphogenesis of human adenovirus type 2: Sequence of entry of proteins into previral and viral particles

Virology ◽

10.1016/0042-6822(84)90256-3 ◽

1984 ◽

Vol 136 (1) ◽

pp. 153-167 ◽

Cited By ~ 14

Author(s):

Nathalie Morin ◽

Pierre Boulanger

Keyword(s):

Human Adenovirus ◽

Adenovirus Type ◽

Viral Particles ◽

Adenovirus Type 2

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Experimental Oral Administration of Canine Adenovirus (Type 2) to Raccoons (Procyon lotor)

Veterinary Pathology ◽

10.1177/030098589202900604 ◽

1992 ◽

Vol 29 (6) ◽

pp. 509-513 ◽

Cited By ~ 10

Author(s):

A. N. Hamir ◽

N. Raju ◽

C. E. Rupprecht

Keyword(s):

Post Infection ◽

Adenovirus Type ◽

Genetically Engineered ◽

Recombinant Vaccines ◽

Intranuclear Inclusions ◽

Ultrastructural Examination ◽

Pulmonary Lesions ◽

Viral Particles ◽

Adenovirus Type 2

Canine adenovirus type 2 (CAV2) has been proposed for recombinant vaccines to control rabies in wild animals. To evaluate the suitability of CAV2 as a safe vector for the genetically engineered vaccines, seven wild-caught raccoons (three males and four females) were administered CAV2 per os. Two of the animals were euthanatized on each of post-infection days 3, 6, and 14, and one was euthanatized on day 21. Two other control raccoons (a male and a female) were also euthanatized on day 21. Microscopic pulmonary lesions of multifocal necrotizing bronchiolitis with basophilic intranuclear inclusions were seen in 3/4 raccoons euthanatized on post-infection days 3 and 6. Ultrastructural examination of lungs with pulmonary lesions revealed hexagonal viral particles characteristic of adenoviruses. CAV2 is potentially pathogenic for raccoons, and this susceptibility should be of concern to developers of recombinant vaccines who intend to use CAV2 as a vaccine vector.

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