scholarly journals Human Adenovirus Type 5 Infection Leads to Nuclear Envelope Destabilization and Membrane Permeability Independently of Adenovirus Death Protein

2021 ◽  
Vol 22 (23) ◽  
pp. 13034
Author(s):  
Søren Pfitzner ◽  
Jens B. Bosse ◽  
Helga Hofmann-Sieber ◽  
Felix Flomm ◽  
Rudolph Reimer ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Nuclear Envelope ◽  
Membrane Permeability ◽  
Nuclear Membrane ◽  
Membrane Damage ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Lamin A ◽  
Infected Cells ◽  
Adenovirus Type 5

The human adenovirus type 5 (HAdV5) infects epithelial cells of the upper and lower respiratory tract. The virus causes lysis of infected cells and thus enables spread of progeny virions to neighboring cells for the next round of infection. The mechanism of adenovirus virion egress across the nuclear barrier is not known. The human adenovirus death protein (ADP) facilitates the release of virions from infected cells and has been hypothesized to cause membrane damage. Here, we set out to answer whether ADP does indeed increase nuclear membrane damage. We analyzed the nuclear envelope morphology using a combination of fluorescence and state-of-the-art electron microscopy techniques, including serial block-face scanning electron microscopy and electron cryo-tomography of focused ion beam-milled cells. We report multiple destabilization phenotypes of the nuclear envelope in HAdV5 infection. These include reduction of lamin A/C at the nuclear envelope, large-scale membrane invaginations, alterations in double membrane separation distance and small-scale membrane protrusions. Additionally, we measured increased nuclear membrane permeability and detected nuclear envelope lesions under cryoconditions. Unexpectedly, and in contrast to previous hypotheses, ADP did not have an effect on lamin A/C reduction or nuclear permeability.

scholarly journals Antiviral Activity of Exopolysaccharides Produced by Lactic Acid Bacteria of the Genera Pediococcus, Leuconostoc and Lactobacillus against Human Adenovirus Type 5

Medicina ◽  
2019 ◽  
Vol 55 (9) ◽  
pp. 519 ◽  
Author(s):  
Biliavska ◽  
Pankivska ◽  
Povnitsa ◽  
Zagorodnya
Keyword(s):  
Cell Cycle ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Flow Cytometric Analysis ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Infected Cells ◽  
Adenovirus Type 5 ◽  
Number Of Cells

Background and objectives: The use of antagonistic probiotic microorganisms and their byproducts represents a promising approach for the treatment of viral diseases. In the current work, the effect of exopolysaccharides (EPSs) produced by lactic acid bacteria from different genera on the structural and functional characteristics of cells and the development of adenoviral infection in vitro was studied. Materials and Methods: Cytotoxicity of six EPSs of lactic acid bacteria of the genera Lactobacillus, Leuconostoc and Pediococcus was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The influence of the EPSs on the infectivity of human adenovirus type 5 (HAdV-5) and on the cell cycle under a condition of adenovirus infection was studied using plaque reduction assay and flow cytometric analysis, respectively. Results: It was shown that exopolysaccharides were non-toxic to Madin-Darby bovine kidney cells (MDBK) as they reduced their viability by 3–17%. A change in the distribution of the cell cycle phases in the non-infected cell population treated with EPSs was observed. The analysis demonstrated an increase in the number of cells in the S phase by 47% when using EPSs 15a and a decrease in the number of cells in the G1 phase by 20–27% when treated with the EPSs 15a, 33a, and 19s. The use of EPSs did not led to the normalization of the life cycle of HAdV-5 infected cells to the level of non-infected cells. The EPSs showed low virucidal activity and reduced the HAdV-5 infectivity to 85%. Among the studied exopolysaccharides, anti-adenovirus activity was found for EPS 26a that is produced by Lactobacillus spp. strain. The treatment of cells with the EPS following virus adsorption completely (100%) suppressed the formation and release of HAdV-5 infectious. Conclusions: EPS 26a possessed distinct anti-HAdV-5 activity and the obtained data demonstrate the potential of using exopolysaccharides as anti-adenoviral agents.

scholarly journals Control of adenovirus early gene expression: posttranscriptional control mediated by both viral and cellular gene products.

1981 ◽  
Vol 1 (9) ◽  
pp. 807-813 ◽  
Author(s):  
M G Katze ◽  
H Persson ◽  
L Philipson
Keyword(s):  
Gene Product ◽  
Messenger Rna ◽  
Adenovirus Type ◽  
Negative Regulator ◽  
Early Gene ◽  
In Vitro Translation ◽  
Mapping Technique ◽  
Cellular Gene ◽  
Infected Cells ◽  
Adenovirus Type 5

An adenovirus type 5 host range mutant (hr-1) located in region E1A and phenotypically defective in expressing viral messenger ribonucleic acid (RNA) from other early regions (Berk et al., Cell 17:935-944, 1979) was analyzed for accumulation of viral RNA in the presence of protein synthesis inhibitors. Nuclear RNA was transcribed from all early regions at the same rate, regardless of whether the drug was present or absent. As expected, low or undetectable levels of RNA were found in the cytoplasm of hr-1-infected cells compared with the wild-type adenovirus type 5 in the absence of drug. When anisomycin was added 30 min before hr-1 infection, cytoplasmic RNA was abundant from early regions E3 and E4 when assayed by filter hybridization. In accordance, early regions E3 and E4 viral messenger RNA species were detected by the S1 endonuclease mapping technique only in hr-1-infected cells that were treated with the drug. Similar results were obtained by in vitro translation studies. Together, these results suggest that this adenovirus type 5 mutant lacks a viral gene product necessary for accumulation of viral messenger RNA, but not for transcription. It is proposed that a cellular gene product serves as a negative regulator of viral messenger RNA accumulation at the posttranscriptional level.

scholarly journals In Vitro and In Vivo Evaluation of Human Adenovirus Type 49 as a Vector for Therapeutic Applications

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1483
Author(s):  
Emily A. Bates ◽  
John R. Counsell ◽  
Sophie Alizert ◽  
Alexander T. Baker ◽  
Natalie Suff ◽  
...  
Keyword(s):  
Viral Vector ◽  
Ex Vivo ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Cell Types ◽  
In Vivo Evaluation ◽  
In Vivo Biodistribution ◽  
Adenovirus Type 5

The human adenovirus phylogenetic tree is split across seven species (A–G). Species D adenoviruses offer potential advantages for gene therapy applications, with low rates of pre-existing immunity detected across screened populations. However, many aspects of the basic virology of species D—such as their cellular tropism, receptor usage, and in vivo biodistribution profile—remain unknown. Here, we have characterized human adenovirus type 49 (HAdV-D49)—a relatively understudied species D member. We report that HAdV-D49 does not appear to use a single pathway to gain cell entry, but appears able to interact with various surface molecules for entry. As such, HAdV-D49 can transduce a broad range of cell types in vitro, with variable engagement of blood coagulation FX. Interestingly, when comparing in vivo biodistribution to adenovirus type 5, HAdV-D49 vectors show reduced liver targeting, whilst maintaining transduction of lung and spleen. Overall, this presents HAdV-D49 as a robust viral vector platform for ex vivo manipulation of human cells, and for in vivo applications where the therapeutic goal is to target the lung or gain access to immune cells in the spleen, whilst avoiding liver interactions, such as intravascular vaccine applications.

Genomic footprinting detects factors bound to major late and IVa2 promoters in adenovirus-infected HeLa cells

1988 ◽  
Vol 8 (4) ◽  
pp. 1534-1539
Author(s):  
G Albrecht ◽  
B Devaux ◽  
C Kedinger
Keyword(s):  
Protein Interactions ◽  
Adenovirus Type ◽  
Nuclease Activity ◽  
Dnase I ◽  
Promoter Activation ◽  
Promoter Elements ◽  
Dnase I Footprinting ◽  
Infected Cells ◽  
Adenovirus Type 5

We used DNase I footprinting assays on nuclei isolated from adenovirus-infected cells to examine the nucleoprotein configuration of a 250-base-pair segment which encompasses the adenovirus type 5 major late (ML) and IVa2 promoters. At 12 and 20 h postinfection (p.i.), fine DNase I digestion mapping of wild-type adenovirus-infected cells revealed specific sequences protected from digestion which corresponded to promoter elements required for expression of the ML gene in vivo. At 12 h p.i., a G+C-rich region which lies upstream of the IVa2 cap site and is important for maximal IVa2 activity was also found masked to nuclease activity. At 20 h p.i., however, this element became more sensitive to nuclease attack, while the ML promoter elements stayed protected. No major changes in DNA-protein interactions were detected in the region spanning the ML and IVa2 cap sites upon promoter activation, suggesting that the binding properties of the cognate factors for this region are not modified during the process.

The endosomal epsilon-coatomer protein is involved in human adenovirus type 5 internalisation

2002 ◽  
Vol 50 (4) ◽  
pp. 481-489 ◽  
Author(s):  
Cs. Jeney ◽  
Boglárka Banizs ◽  
Orsolya Dobay ◽  
Keyword(s):  
Chinese Hamster ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Inhibitory Effect ◽  
Bafilomycin A1 ◽  
Cho Cell ◽  
Coxsackie Adenovirus Receptor ◽  
Downstream Effector ◽  
Adenovirus Receptor ◽  
Adenovirus Type 5

The effects of bafilomycin A1 and of the reduced level of endosomal epsilon-COP (coatomer protein) on the infectivity of human adenovirus type 5 were investigated in Coxsackie adenovirus receptor- (CAR-) transfected Chinese hamster ovary (CHO) cells. The endosomal proton pump inhibitor bafilomycin A1 was able to cause only partial inhibition. Using ldlF cells (an epsilon-COP thermosensitive mutant CHO cell line) the reduction of epsilon-COP level also had partial inhibitory effect. Based on these results and comparing them to existing models of the adenovirus entry, we propose a refined model in which there are two pathways of adenoviral entry: the first one involves the epsilon-COP as the downstream effector of the acidification and can be blocked by bafilomycin A1 and the second one is a pH-independent pathway.

scholarly journals Chimpanzee adenovirus CV-68 adapted as a gene delivery vector interacts with the coxsackievirus and adenovirus receptor

2002 ◽  
Vol 83 (1) ◽  
pp. 151-155 ◽  
Author(s):  
Christopher J. Cohen ◽  
Zhi Quan Xiang ◽  
Guang-Ping Gao ◽  
Hildegund C. J. Ertl ◽  
James M. Wilson ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Cho Cells ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Soluble Form ◽  
Delivery Vector ◽  
Coxsackievirus And Adenovirus Receptor ◽  
Adenovirus Receptor ◽  
Adenovirus Type 5 ◽  
Dependent Mechanism

A replication-defective form of chimpanzee adenovirus type 68 (C68) has been developed to circumvent problems posed by widespread preexisting immunity to common human adenovirus vectors. To investigate the determinants of C68 tropism, its interaction with the coxsackievirus and adenovirus receptor (CAR) was studied. Although CHO cells were resistant to transduction by C68 as well as by adenovirus type 5 (Ad5), CHO cells expressing either human or murine CAR were transduced readily. C68 transduction, like Ad5 transduction, was blocked when cells were exposed to anti-CAR antibody or when virus was exposed to a soluble form of the CAR extracellular domain. These results indicate that gene delivery by C68 occurs by a CAR-dependent mechanism.

scholarly journals Bovine herpesvirus 1 glycoprotein D expression in bovine upper respiratory tract mediated by a human adenovirus type 5

2004 ◽  
Vol 35 (6) ◽  
pp. 715-721 ◽  
Author(s):  
Sacha Gogev ◽  
Jean-Pierre Georgin ◽  
Fr�d�ric Schynts ◽  
Alain Vanderplasschen ◽  
Etienne Thiry
Keyword(s):  
Respiratory Tract ◽  
Bovine Herpesvirus ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Upper Respiratory Tract ◽  
Glycoprotein D ◽  
Bovine Herpesvirus 1 ◽  
Upper Respiratory ◽  
Herpesvirus 1 ◽  
Adenovirus Type 5

scholarly journals Comparative Sequence Analysis of the Largest E1A Proteins of Human and Simian Adenoviruses

2002 ◽  
Vol 76 (16) ◽  
pp. 7968-7975 ◽  
Author(s):  
Nikita Avvakumov ◽  
Russ Wheeler ◽  
Jean Claude D'Halluin ◽  
Joe S. Mymryk
Keyword(s):  
Gene Expression ◽  
Human Adenovirus ◽  
Detailed Comparison ◽  
Adenovirus Type ◽  
Comparative Sequence Analysis ◽  
Regulatory Proteins ◽  
Carboxyl Terminus ◽  
Adenovirus E1a ◽  
Simian Adenovirus ◽  
Adenovirus Type 5

ABSTRACT The early region 1A (E1A) gene is the first gene expressed after infection with adenovirus and has been most extensively characterized in human adenovirus type 5 (hAd5). The E1A proteins interact with numerous cellular regulatory proteins, influencing a variety of transcriptional and cell cycle events. For this reason, these multifunctional proteins have been useful as tools for dissecting pathways regulating cell growth and gene expression. Despite the large number of studies using hAd5 E1A, relatively little is known about the function of the E1A proteins of other adenoviruses. In 1985, a comparison of E1A sequences from three human and one simian adenovirus identified three regions with higher overall levels of sequence conservation designated conserved regions (CR) 1, 2, and 3. As expected, these regions are critical for a variety of E1A functions. Since that time, the sequences of several other human and simian adenovirus E1A proteins have been determined. Using these, and two additional sequences that we determined, we report here a detailed comparison of the sequences of 15 E1A proteins representing each of the six hAd subgroups and several simian adenoviruses. These analyses refine the positioning of CR1, 2, and 3; define a fourth CR located near the carboxyl terminus of E1A; and suggest several new functions for E1A.

Further observations on nucleolar tails in amphibian oocytes

1991 ◽  
Vol 99 (3) ◽  
pp. 515-521
Author(s):  
PEDRO LEÓN ◽  
JAMES KEZER ◽  
ERIC SCHABTACH
Keyword(s):  
Electron Microscopy ◽  
Nuclear Envelope ◽  
Nuclear Membrane ◽  
Outer Surface ◽  
Light And Electron Microscopy ◽  
Nuclear Pores ◽  
Core Component ◽  
Amphibian Species ◽  
Attachment Structures ◽  
Convoluted Tubules

Large oocytes from some amphibian species possess beaded or unbeaded intranuclear tails that penetrate the extrachromosomal nucleoli through a distinct pit in their surface and attach to the central core component Here we show, using light and electron microscopy, that tails anchor nucleoli to the nuclear envelope through intricate attachment structures. These structures are composed of interconnected spherical masses containing highly convoluted tubules and associated extratubular proteins, directly directly in contact with the inner nuclear membrane. Fibers emerging from the nuclear pores seemingly hold the attachment complex in place. Beads on the nucleolar tails are formed by the accumulation of proteins on the outer surface of smooth tubules. The function of these intranuclear tubules is unknown

Mapping of cellular protein-binding sites on the products of early-region 1A of human adenovirus type 5

1988 ◽  
Vol 8 (9) ◽  
pp. 3955-3959 ◽  
Author(s):  
C Egan ◽  
T N Jelsma ◽  
J A Howe ◽  
S T Bayley ◽  
B Ferguson ◽  
...  
Keyword(s):  
Biological Activity ◽  
Binding Sites ◽  
Human Adenovirus ◽  
Adenovirus Type ◽  
Cellular Protein ◽  
Cellular Proteins ◽  
Deletion Mutants ◽  
Protein Binding Sites ◽  
Amino Terminal ◽  
Adenovirus Type 5

The binding sites for the 300-, 107-, and 105-kilodalton cellular proteins which associate with human adenovirus type 5 E1A products were studied with E1A deletion mutants. All appeared to bind to the amino-terminal half of E1A products in regions necessary for oncogenic transformation. These results suggest that these cellular species may be important for the biological activity of E1A products.

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