Proteolytic Cleavage of Bovine Adenovirus 3-Encoded pVIII

Gaba, Amit; Ayalew, Lisanework E.; Makadiya, Niraj; Tikoo, Suresh K.

doi:10.1128/jvi.00211-17

Cited by 7 publications

(8 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Unlike BAV304a, electron microcopic analysis detected disrupted virions in CsCl gradient purified mutant virus. Earlier, we reported that the cleaved C‐terminus (amino acid 147–216) of proteinVIII interacts with hexon, which is essential for the stability of BAdV‐3 virion (Ayalew et al, ; Gaba, Ayalew, Makadiya, & Tikoo, ). Thus, the deletion of amino acids 147–174 may affect the interaction of protein VIII with hexon, required for the stability of mature BAdV‐3 virion leading to the production of unstable virions, with disrupted capsids due to exposure to CsCl/TEM procedure.…”

Section: Discussionmentioning

confidence: 99%

Bovine adenovirus‐3 protein VIII associates with eukaryotic initiation factor‐6 during infection

Gaba

Ayalew

Patel

et al. 2018

Cellular Microbiology

Self Cite

View full text Add to dashboard Cite

Adenovirus protein VIII appears to connect core with the inner surface of the adenovirus capsid. Because protein-protein interactions are central to virus replication, identification of proteins interacting with protein VIII may help in understanding their role in adenovirus infection. Our yeast 2-hybrid assay indicated that protein VIII interacts with eukaryotic initiation factor 6 (eIF6). These findings were confirmed by Glutathione S-transferase-pull down assay, bimolecular fluorescent complementation assay, and coimmunoprecipitation assay in plasmid DNA transfected and bovine adenovirus-3 (BAdV-3) infected cells. The C-terminus amino acids 147 to 174 of protein VIII and N-terminus amino acids 44 to 97 of eIF6 are involved in these interactions. Polysome analysis demonstrated increased level of 60S ribosomal subunit and decreased level of 80S complex in protein VIII expressing cells or BAdV-3 infected cells. Our results suggest that formation of functional 80S ribosome appears impaired in the presence of protein VIII at late times post infection. We speculate that this impaired ribosome assembly may be responsible for the inhibition of cellular mRNA translation observed late in adenovirus infected cells. Moreover, analysis of recombinant BAdV-3 expressing mutant protein VIII (deletion of eIF6 interacting domain) suggests that interaction of protein VIII and eIF6 may help in preferential translation of adenovirus genes during late phase of adenovirus infection.

show abstract

Section: Discussionmentioning

confidence: 99%

Bovine adenovirus‐3 protein VIII associates with eukaryotic initiation factor‐6 during infection

Gaba

Ayalew

Patel

et al. 2018

Cellular Microbiology

Self Cite

View full text Add to dashboard Cite

show abstract

“…The cell pellets were collected, and genomic DNA was extracted. The viral genome copy number was determined by quantitative PCR using primers (Table 3) as described previously (46).…”

Section: Discussionmentioning

confidence: 99%

“…Earlier reports analyzing the atomic models of adenovirus have predicted the association of different structural proteins, which may be required for proper assembly of adenovirus and formation of stable virions (42)(43)(44)(45). Further analysis of mutant adenoviruses suggested that deletion/mutation of adenovirus proteins, including IVa2 (34), VIII (46), and pV (40,47), leads to the production of disrupted/aberrant capsids. Our results demonstrate that inactivation of E3 13.7K protein expression results in the production of disfigured capsids, suggesting that 13.7K appears to be essential for maintaining the stability of PAdV-3 virions.…”

Section: Discussionmentioning

confidence: 99%

“…VIDO R1 cells (6) infected with PAdV-3 or PAV13.7 3A were harvested at indicated times postinfection. After centrifugation, the cell pellets were collected, genomic DNA was extracted, and the viral genome copy number was determined by quantitative real-time PCR as described previously (46). Specifically, two sets of oligonucleotide primers (␤-actin FW and ␤-actin RV; Hexon FW and Hexon RV), listed in Table 3, were used in quantitative PCR.…”

Section: Construction Of Vido As2 Cells (Vido R1 Cell Line Expressingmentioning

confidence: 99%

See 1 more Smart Citation

Porcine Adenovirus Type 3 E3 Encodes a Structural Protein Essential for Capsid Stability and Production of Infectious Progeny Virions

Said

Woldemariam

Tikoo

2018

J Virol

View full text Add to dashboard Cite

The adenovirus E3 region encodes proteins that are not essential for viral replication The porcine adenovirus type 3 (PAdV-3) E3 region encodes three proteins, including 13.7K. Here, we report that 13.7K is expressed as an early protein, which localizes to the nucleus of infected cells. The 13.7K protein is a structural protein, as it is incorporated in CsCl-purified virions. The 13.7K protein appears to be essential for PAdV-3 replication, as mutant PAV13.7 expressing a mutated 13.7K could be isolated only in VIDO AS2 cells expressing the 13.7K protein. Analysis of PAV13.7 suggested that even in the presence of reduced levels of some late viral proteins, there appeared to be no effect on virus assembly and production of mature virions. Further analysis of CsCl-purified PAV13.7 by transmission electron microscopy revealed the presence of disrupted/broken capsids, suggesting that inactivation of 13.7K protein expression may produce fragile capsids. Our results suggest that the PAdV-3 E3 region-encoded 13.7K protein is a capsid protein, which appears to be essential for the formation of stable capsids and production of infectious progeny virions. Although E3 region-encoded proteins are involved in the modulation of leukocyte functions (N. Arnberg, Proc Natl Acad Sci U S A 110:19976-19977, 2013) and inducing a lytic infection of lymphocytes (V. K. Murali, D. A. Ornelles, L. R. Gooding, H. T. Wilms, W. Huang, A. E. Tollefson, W. S. Wold, and C. Garnett-Benson, J Virol 88:903-912, 2014), none of the E3 proteins appear to be a component of virion capsid or required for replication of adenovirus. Here, we demonstrate that the 13.7K protein encoded by the E3 region of porcine adenovirus type 3 is a component of progeny virion capsids and appears to be essential for maintaining the integrity of virion capsid and production of infectious progeny virions. To our knowledge, this is the first report to suggest that an adenovirus E3-encoded protein is an essential structural protein.

show abstract

“…At 48 h post-infection, the cells were harvested. The virus infected cells of CsCl-purified BAdV-3 virions were processed as described earlier ( Zhao and Tikoo, 2016 ; Gaba et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

Nuclear and Nucleolar Localization of Bovine Adenovirus-3 Protein V

Zhao

Tikoo

2021

Front. Microbiol.

View full text Add to dashboard Cite

The L2 region of bovine adenovirus-3 (BAdV-3) encodes a Mastadenovirus genus-specific protein, designated as pV, which is important for the production of progeny viruses. Here, we demonstrate that BAdV-3 pV, expressed as 55 kDa protein, localizes to the nucleus and specifically targets nucleolus of the infected cells. Analysis of deletion mutants of pV suggested that amino acids 81–120, 190–210, and 380–389 act as multiple nuclear localization signals (NLS), which also appear to serve as the binding sites for importin α-3 protein, a member of the importin α/β nuclear import receptor pathway. Moreover, pV amino acids 21–50 and 380–389 appear to act as nucleolar localization signals (NoLs). Interestingly, amino acids 380–389 appear to act both as NLS and as NoLS. The presence of NoLS is essential for the production of infectious progeny virions, as deletion of both NoLs are lethal for the production of infectious BAdV-3. Analysis of mutant BAV.pVd1d3 (isolated in pV completing CRL cells) containing deletion/mutation of both NoLS in non-complementing CRL cells not only revealed the altered intracellular localization of mutant pV but also reduced the expression of some late proteins. However, it does not appear to affect the incorporation of viral proteins, including mutant pV, in BAV.pVd1d3 virions. Further analysis of CsCl purified BAV.pVd1d3 suggested the presence of thermo-labile virions with disrupted capsids, which appear to affect the infectivity of the progeny virions. Our results suggest that pV contains overlapping and non-overlapping NoLS/NLS. Moreover, the presence of both NoLS appear essential for the production of stable and infectious progeny BAV.pVd1d3 virions.

show abstract

Proteolytic Cleavage of Bovine Adenovirus 3-Encoded pVIII

Cited by 7 publications

References 43 publications

Bovine adenovirus‐3 protein VIII associates with eukaryotic initiation factor‐6 during infection

Bovine adenovirus‐3 protein VIII associates with eukaryotic initiation factor‐6 during infection

Porcine Adenovirus Type 3 E3 Encodes a Structural Protein Essential for Capsid Stability and Production of Infectious Progeny Virions

Nuclear and Nucleolar Localization of Bovine Adenovirus-3 Protein V

Contact Info

Product

Resources

About