Effects of tunicamycin and monensin on the distribution of highly phosphorylated proteins in cells infected with herpes simplex virus type 1

López‐Iglesias, Carmen; Puvion‐Dutilleul, Francine

doi:10.1016/0889-1605(88)90007-9

Cited by 4 publications

(3 citation statements)

References 23 publications

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“…After being concentrated within the rough endoplamic reticulum and Golgi apparatus, ADP migrate to the nuclear envelope of cells at 33 to 41 post-infection (Scaria et al, 1992). Roughly similar changes were observed in monensin-treated herpes simplex virus type 1 infected cells, a drug which blocks the migration of Golgi apparatus vesicles (Lopez-Iglesias and Puvion-Dutilleul, 1988). It cannot be excluded that ADP might alter the nuclear envelope rendering it locally weak.…”

Section: Discussionmentioning

confidence: 66%

Release of viruses and viral DNA from nucleus to cytoplasm of HeLa cells at late stages of productive adenovirus infection as revealed by electron microscope in situ hybridization

Puvion‐Dutilleul¹,

Besse

Pichard³

et al. 1998

Biology of the Cell

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Considerable progress has been made over the past 10 years towards a full understanding of the functional significance of the structural changes resulting from the production of adenoviruses in permissive cells. Similarly, the host-virus interactions which are involved in viral replication and gene expression as well as in RNA nuclear export have been investigated. Post-embedding nonisotopic in situ hybridization has been proven to be a powerful tool for the study of nucleic acids in infected cells provided that controlled elimination of artifacts by appropriate treatments was undertaken. Adenovirus infected cells present two biological characteristics which could lead to false positive or negative results. First, they contain large amounts of single-stranded portions of viral DNA which are revealed with viral RNA molecules. Second, DNA-binding proteins are present which hide some nucleic acid sequences. By using a DNA probe and appropriate variations in the experimental protocol, it is possible to reveal specifically different kinds of targets, simultaneously single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), or only ssDNA, or only dsDNA, or only RNA. By using two probes labeled with different haptens, biotine and digoxigenin, it is possible to detect concomitantly two nucleic acid targets and, therefore, to study their relationships. In order to gain insight into the changes in the nucleus before cell lysis and to improve our knowledge on the series of steps leading to the release of adenoviruses from the nucleus, examination of cells at 41 h post-infection and identification of structures containing adenoviral nucleic acids were undertaken. In addition to the ultrastructural changes and precise distribution of cellular DNA and viral nucleic acid molecules already described in cells up to 24 h post-infection (for a review, see Puvion E, Puvion-Dutilleul F (1996) Exp Cell Res 229, 217-225), new results were obtained. Routine observation revealed the presence of: i) viruses in the cytoplasm, some being located next to nuclear pores; ii) abnormally large portions of the nuclear envelope devoid of underlying condensed chromatin; iii) proliferation of either the inner nuclear membrane only or both membranes of the nuclear envelope; and iv) electron-opaque grains in the nuclear compartment involved in viral genome transcription, and also in the clusters of interchromatin granules known to contain mature viral messenger RNA (Bridge E et al (1996) J Cell Biol 135, 303-314). In situ hybridization revealed the presence of: i) dsDNA in the cytoplasmic viruses indicating that they were mature viruses; ii) free viral dsDNA and ssDNA molecules in the cytoplasm whereas host DNA remained confined at the nuclear border; and iii) viral RNA in the newly-described electron-opaque grains we call, therefore, viral-RNA containing grains. Immunodetection of bromodeoxyuridine (BrdU) incorporated into DNA in pulse and pulse-chase experiments allowed us to ascertain that cells at 41 h post-infection were truly living cells and ...

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Section: Discussionmentioning

confidence: 66%

Release of viruses and viral DNA from nucleus to cytoplasm of HeLa cells at late stages of productive adenovirus infection as revealed by electron microscope in situ hybridization

Puvion‐Dutilleul¹,

Besse

Pichard³

et al. 1998

Biology of the Cell

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“…Nigericin and salinomycin inhibited Wnt signaling by blocking the phosphorylation of the Wnt coreceptor LRP6 and inducing its degradation (12,(31)(32)(33)(34). Monensin was reported in the past to block protein transport from the Golgi apparatus to the cell membrane and to inhibit HSV-1, HSV-2, and HCMV (35)(36)(37)(38). Monensin treatment inhibited transport of progeny virus to the surface of infected cells, while viral protein synthesis and DNA replication were not inhibited.…”

Section: Discussionmentioning

confidence: 99%

Wnt Modulating Agents Inhibit Human Cytomegalovirus Replication

Kapoor

Venkatadri

et al. 2013

Antimicrob Agents Chemother

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“…After 20 minutes, cells were scraped and transferred to a 15 ml flask, and pelleted at 1,500 rpm for five minutes. Pellets were fixed and included in resin according to standard procedures [71]–[73]. Electron microscopy images were obtained using a Tecnai Spirit Twin (FEI, Hillsboro, OR) and processed using ImageJ [http://rsbweb.nih.gov/ij/index.html].…”

Section: Methodsmentioning

confidence: 99%

Generation and Characterization of a Defective HIV-1 Virus as an Immunogen for a Therapeutic Vaccine

et al. 2012

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BackgroundThe generation of new immunogens able to elicit strong specific immune responses remains a major challenge in the attempts to obtain a prophylactic or therapeutic vaccine against HIV/AIDS. We designed and constructed a defective recombinant virus based on the HIV-1 genome generating infective but non-replicative virions able to elicit broad and strong cellular immune responses in HIV-1 seropositive individuals.ResultsViral particles were generated through transient transfection in producer cells (293-T) of a full length HIV-1 DNA carrying a deletion of 892 base pairs (bp) in the pol gene encompassing the sequence that codes for the reverse transcriptase (NL4-3/ΔRT clone). The viral particles generated were able to enter target cells, but due to the absence of reverse transcriptase no replication was detected. The immunogenic capacity of these particles was assessed by ELISPOT to determine γ-interferon production in a cohort of 69 chronic asymptomatic HIV-1 seropositive individuals. Surprisingly, defective particles produced from NL4-3/ΔRT triggered stronger cellular responses than wild-type HIV-1 viruses inactivated with Aldrithiol-2 (AT-2) and in a larger proportion of individuals (55% versus 23% seropositive individuals tested). Electron microscopy showed that NL4-3/ΔRT virions display immature morphology. Interestingly, wild-type viruses treated with Amprenavir (APV) to induce defective core maturation also induced stronger responses than the same viral particles generated in the absence of protease inhibitors.ConclusionsWe propose that immature HIV-1 virions generated from NL4-3/ΔRT viral clones may represent new prototypes of immunogens with a safer profile and stronger capacity to induce cellular immune responses than wild-type inactivated viral particles.

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Effects of tunicamycin and monensin on the distribution of highly phosphorylated proteins in cells infected with herpes simplex virus type 1

Cited by 4 publications

References 23 publications

Release of viruses and viral DNA from nucleus to cytoplasm of HeLa cells at late stages of productive adenovirus infection as revealed by electron microscope in situ hybridization

Release of viruses and viral DNA from nucleus to cytoplasm of HeLa cells at late stages of productive adenovirus infection as revealed by electron microscope in situ hybridization

Wnt Modulating Agents Inhibit Human Cytomegalovirus Replication

Generation and Characterization of a Defective HIV-1 Virus as an Immunogen for a Therapeutic Vaccine

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