Thermostability/Infectivity Defect Caused by Deletion of the Core Protein V Gene in Human Adenovirus Type 5 Is Rescued by Thermo-selectable Mutations in the Core Protein X Precursor

Ugai, Hideyo; Borovjagin, Anton V.; Le, Long P.; Wang, Minghui; Curiel, David T.

doi:10.1016/j.jmb.2006.11.090

Cited by 32 publications

(50 citation statements)

References 52 publications

(10 reference statements)

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“…A mutant adenovirus that lacks protein V (Ad5-dV) has reduced thermostability and infectivity, demonstrating that protein V plays a critical role in the generation of infectious particles (21). However, relatively little biochemical or structural information has been obtained for protein V. During the infection process, protein V is not released from the viral particles either when the virus penetrates the cells during infection (43) or upon heating purified particles (42).…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 94%

“…This deletion significantly disrupted viral assembly concomitant with alterations in the capsid morphology and a substantial reduction in thermostability and infectivity (21). Overall, these studies indicate that protein V has a critical role in capsid assembly and in the formation of infectious virions (21). Protein V contains multiple nuclear and nucleolar localization signals and is imported into the nucleus very shortly after viral infection, suggesting that it has a role in the delivery of viral DNA during the infection process (20,22).…”

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confidence: 95%

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Isolation and Characterization of the DNA and Protein Binding Activities of Adenovirus Core Protein V

Pérez‐Vargas

Vaughan

Houser

et al. 2014

J Virol

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The structure of adenovirus outer capsid was revealed recently at 3-to 4-Å resolution (V. Reddy, S. Natchiar, P. Stewart, and G. Nemerow, Science 329:1071-1075, 2010, http://dx.doi.org/10.1126/science.1187292); however, precise details on the function and biochemical and structural features for the inner core still are lacking. Protein V is one the most important components of the adenovirus core, as it links the outer capsid via association with protein VI with the inner DNA core. Protein V is a highly basic protein that strongly binds to DNA in a nonspecific manner. We report the expression of a soluble protein V that exists in monomer-dimer equilibrium. Using reversible cross-linking affinity purification in combination with mass spectrometry, we found that protein V contains multiple DNA binding sites. The binding sites from protein V mediate heat-stable nucleic acid associations, with some of the binding sites possibly masked in the virus by other core proteins. We also demonstrate direct interaction between soluble proteins V and VI, thereby revealing the bridging of the inner DNA core with the outer capsid proteins. These findings are consistent with a model of nucleosome-like structures proposed for the adenovirus core and encapsidated DNA. They also suggest an additional role for protein V in linking the inner nucleic acid core with protein VI on the inner capsid shell. IMPORTANCEScant knowledge exists of how the inner core of adenovirus containing its double-stranded DNA (dsDNA) genome and associated proteins is organized. Here, we report a purification scheme for a recombinant form of protein V that allowed analysis of its interactions with the nucleic acid core region. We demonstrate that protein V exhibits stable associations with dsDNA due to the presence of multiple nucleic acid binding sites identified both in the isolated recombinant protein and in virus particles. As protein V also binds to the membrane lytic protein VI molecules, this core protein may serve as a bridge from the inner dsDNA core to the inner capsid shell.

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Section: Discussionmentioning

confidence: 99%

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confidence: 94%

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confidence: 95%

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Isolation and Characterization of the DNA and Protein Binding Activities of Adenovirus Core Protein V

Pérez‐Vargas

Vaughan

Houser

et al. 2014

J Virol

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“…The role of core protein V is less certain, but it was recently shown that it is nonessential for virus viability, although the initial mutants produced were extremely thermolabile. Variants selected for greater thermostability showed mutations in core protein X, suggestive of an interaction between the two proteins (34). One copy of terminal protein is also bound to each end of the genome.…”

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confidence: 99%

Cryoelectron Microscopy Map of Atadenovirus Reveals Cross-Genus Structural Differences from Human Adenovirus

Pantelic

Lockett

Rothnagel

et al. 2008

J Virol

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A three-dimensional (3D) cryoelectron microscopy reconstruction of the prototype Atadenovirus (OAdV [an ovine adenovirus isolate]) showing information at a 10.6-Å resolution (0.5 Fourier shell correlation) was derived by single-particle analysis. This is the first 3D structure solved for any adenovirus that is not a Mastadenovirus, allowing cross-genus comparisons between structures and the assignment of genus-specific capsid proteins. Viable OAdV mutants that lacked the genus-specific LH3 and p32k proteins in purified virions were also generated. Negatively stained 3D reconstructions of these mutants were used to identify the location of protein LH3 and infer that of p32k within the capsid. The key finding was that LH3 is a critical protein that holds the outer capsid of the virus together. In its absence, the outer viral capsid is unstable. LH3 is located in the same position among the hexon subunits as its protein IX equivalent from mastadenoviruses but sits on top of the hexon trimers, forming prominent "knobs" on the virion surface that visually distinguish OAdV from other known AdVs. Electron density was also assigned to hexon and penton subunits and to proteins IIIa and VIII. There was good correspondence between OAdV density and human AdV hexon structures, which also validated the significant differences that were observed between the penton base protein structures.The Adenoviridae comprise a large family of non-enveloped, icosahedral, generally nonpathogenic viruses with a doublestranded DNA genome (ϳ26.1 to 45 kb) (9). Because of their wide host range, viruses such as human adenovirus type 5 (AdV5) have been developed as gene delivery vectors and applied in the clinic, especially for cancer, monogenic disorders, and vaccination (http://www.wiley.co.uk/genmed /clinical/). While some of these trials have reached phases II and III, there is still considerable room for improvement in vector technology. In particular, there is strong interest in developing vectors that are targeted to specific tissues for improved therapeutic effect (6, 11), and for this purpose, structural information on the virus particle and its constituent molecules is paramount.The family Adenoviridae comprises four genera (Mastadenovirus, Aviadenovirus, Siadenovirus, and Atadenovirus). Of these, the best studied are the mastadenoviruses, which include viruses from numerous mammalian species and all known human AdVs. In contrast, members of the avi-, si-, and atadenoviruses have been isolated from birds, although the latter also occur widely in mammalian and reptilian species (9, 38). The prototype Atadenovirus, an ovine isolate (serotype 7; OAdV), is the only member for which any significant biological studies have been undertaken. OAdV uses a different (unknown) receptor from AdV5 and infects, but does not replicate in, human cells (15). It has also been developed as a gene delivery vector (2) and has regulatory approval to enter a phase I clinical trial for prostate cancer in 2008.Representative genomes from all four genera have now...

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“…It is not known if pV enters the nucleus on its own or in complex with other factors (143), or if it stays in the cytoplasm in association with the mitochondrial protein p32 implicated in ARFmediated apoptosis (144,145). Remarkably, a pV knock-out virus is viable in the presence of compensatory mutations in the mu protein (146). This suggests that pV has no unique functions in the replication cycle of mastadenoviruses, at least in cultured cells, which is further supported by the observation that pV does not occur in atadenoviruses, aviadenoviruses and siadenoviruses.…”

Section: Nuclear Import Of Adenovirus Dna Associated Proteinsmentioning

confidence: 99%

DNA-tumor virus entry—From plasma membrane to the nucleus

Greber

Püntener

2009

Seminars in Cell & Developmental Biology

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DNA-tumor viruses comprise enveloped and non-enveloped agents that cause malignancies in a large variety of cell types and tissues by interfering with cell cycle control and immortalization. Those DNA-tumor viruses that replicate in the nucleus use cellular mechanisms to transport their genome and newly synthesized viral proteins into the nucleus. This requires cytoplasmic transport and nuclear import of their genome. Agents that employ this strategy include adenoviruses, hepadnaviruses, herpesviruses, and likely also papillomaviruses, and polyomaviruses, but not poxviruses which replicate in the cytoplasm. Here, we discuss how DNA-tumor viruses enter cells, take advantage of cytoplasmic transport, and import their DNA genome through the nuclear pore complex into the nucleus. Remarkably, nuclear import of incoming genomes does not necessarily follow the same pathways used by the structural proteins of the viruses during the replication and assembly phases of the viral life cycle. Understanding the mechanisms of DNA nuclear import can identify new pathways of cell regulation and anti-viral therapies.

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Thermostability/Infectivity Defect Caused by Deletion of the Core Protein V Gene in Human Adenovirus Type 5 Is Rescued by Thermo-selectable Mutations in the Core Protein X Precursor

Cited by 32 publications

References 52 publications

Isolation and Characterization of the DNA and Protein Binding Activities of Adenovirus Core Protein V

Isolation and Characterization of the DNA and Protein Binding Activities of Adenovirus Core Protein V

Cryoelectron Microscopy Map of Atadenovirus Reveals Cross-Genus Structural Differences from Human Adenovirus

DNA-tumor virus entry—From plasma membrane to the nucleus

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