scholarly journals Chikungunya virus requires an intact microtubule network for efficient viral genome delivery

2020 ◽  
Author(s):  
Tabitha E. Hoornweg ◽  
Ellen M. Bouma ◽  
Denise P.I. van de Pol ◽  
Izabela A. Rodenhuis-Zybert ◽  
Jolanda M. Smit
Keyword(s):  
Host Cell ◽  
Viral Genome ◽  
Intracellular Trafficking ◽  
Chikungunya Virus ◽  
Antiviral Treatment ◽  
Cell Entry ◽  
Cell Periphery ◽  
Microtubule Network ◽  
Enveloped Virus ◽  
Early Endosomes

AbstractChikungunya virus (CHIKV) is a re-emerging mosquito-borne virus, which has rapidly spread around the globe thereby causing millions of infections. CHIKV is an enveloped virus belonging to the Togaviridae family and enters its host cell primarily via clathrin-mediated endocytosis. Upon internalization, the endocytic vesicle containing the virus particle moves through the cell and delivers the virus to early endosomes where membrane fusion is observed. Thereafter, the nucleocapsid dissociates and the viral RNA is translated into proteins. In this study, we examined the importance of the microtubule network during the early steps of infection and dissected the intracellular trafficking behavior of CHIKV particles during cell entry. We observed two distinct CHIKV intracellular trafficking patterns prior to membrane hemifusion. Whereas half of the CHIKV virions remained static during cell entry and fused in the cell periphery, the other half showed fast-directed microtubule-dependent movement prior to delivery to Rab5-positive early endosomes and predominantly fused in the perinuclear region of the cell. Disruption of the microtubule network reduced the number of infected cells. At these conditions, membrane hemifusion activity was not affected yet fusion was restricted to the cell periphery. Furthermore, follow-up experiments revealed that disruption of the microtubule network impairs the delivery of the viral genome to the cell cytosol. We therefore hypothesize that microtubules may direct the particle to a cellular location that is beneficial for establishing infection or aids in nucleocapsid uncoating.Author SummaryChikungunya virus (CHIKV) is an alphavirus that is transmitted to humans by infected mosquitoes. Disease symptoms can include fever, rash, myalgia, and long-lasting debilitating joint pains. Unfortunately, there is currently no licensed vaccine or antiviral treatment available to combat CHIKV. Understanding the virus:host interactions during the replication cycle of the virus is crucial for the development of effective antiviral therapies. In this study we elucidated the trafficking behavior of CHIKV particles early in infection. During cell entry, CHIKV virions require an intact microtubule network for efficient delivery of the viral genome into the host cell thereby increasing the chance to productively infect a cell.

scholarly journals Dynamics of Chikungunya Virus Cell Entry Unraveled by Single-Virus Tracking in Living Cells

2016 ◽  
Vol 90 (9) ◽  
pp. 4745-4756 ◽  
Author(s):  
Tabitha E. Hoornweg ◽  
Mareike K. S. van Duijl-Richter ◽  
Nilda V. Ayala Nuñez ◽  
Irina C. Albulescu ◽  
Martijn J. van Hemert ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Chikungunya Virus ◽  
Antiviral Treatment ◽  
Febrile Illness ◽  
Living Cells ◽  
Cell Entry ◽  
Entry Pathway ◽  
Early Endosomes ◽  
Entry Behavior ◽  
Insight Into

ABSTRACTChikungunya virus (CHIKV) is a rapidly emerging mosquito-borne human pathogen causing major outbreaks in Africa, Asia, and the Americas. The cell entry pathway hijacked by CHIKV to infect a cell has been studied previously using inhibitory compounds. There has been some debate on the mechanism by which CHIKV enters the cell: several studies suggest that CHIKV enters via clathrin-mediated endocytosis, while others show that it enters independently of clathrin. Here we applied live-cell microscopy and monitored the cell entry behavior of single CHIKV particles in living cells transfected with fluorescent marker proteins. This approach allowed us to obtain detailed insight into the dynamic events that occur during CHIKV entry. We observed that almost all particles fused within 20 min after addition to the cells. Of the particles that fused, the vast majority first colocalized with clathrin. The average time from initial colocalization with clathrin to the moment of membrane fusion was 1.7 min, highlighting the rapidity of the cell entry process of CHIKV. Furthermore, these results show that the virus spends a relatively long time searching for a receptor. Membrane fusion was observed predominantly from within Rab5-positive endosomes and often occurred within 40 s after delivery to endosomes. Furthermore, we confirmed that a valine at position 226 of the E1 protein enhances the cholesterol-dependent membrane fusion properties of CHIKV. To conclude, our work confirms that CHIKV enters cells via clathrin-mediated endocytosis and shows that fusion occurs from within acidic early endosomes.IMPORTANCESince its reemergence in 2004, chikungunya virus (CHIKV) has spread rapidly around the world, leading to millions of infections. CHIKV often causes chikungunya fever, a self-limiting febrile illness with severe arthralgia. Currently, no vaccine or specific antiviral treatment against CHIKV is available. A potential antiviral strategy is to interfere with the cell entry process of the virus. However, conflicting results with regard to the cell entry pathway used by CHIKV have been published. Here we applied a novel technology to visualize the entry behavior of single CHIKV particles in living cells. Our results show that CHIKV cell entry is extremely rapid and occurs via clathrin-mediated endocytosis. Membrane fusion from within acidic early endosomes is observed. Furthermore, the membrane fusion capacity of CHIKV is strongly promoted by cholesterol in the target membrane. Taking these findings together, this study provides detailed insight into the cell entry process of CHIKV.

scholarly journals Intracellular Trafficking of Bordetella pertussis in Human Macrophages

2010 ◽  
Vol 78 (3) ◽  
pp. 907-913 ◽  
Author(s):  
Yanina A. Lamberti ◽  
Jimena Alvarez Hayes ◽  
Maria L. Perez Vidakovics ◽  
Eric T. Harvill ◽  
Maria Eugenia Rodriguez
Keyword(s):  
Host Cell ◽  
Bordetella Pertussis ◽  
Intracellular Trafficking ◽  
Intracellular Bacteria ◽  
Human Macrophages ◽  
Early Endosomes ◽  
Viable Bacteria ◽  
Cell Recycling ◽  
Recycling Pathway ◽  
Acidic Compartments

ABSTRACT Although Bordetella pertussis has been observed to survive inside macrophages, its ability to resist or evade degradation in phagolysosomes has not been defined. We here investigated the trafficking of B. pertussis upon entry into human macrophages. During the first hours following phagocytosis, a high percentage of bacteria were destroyed within acidic compartments positive for the lysosome-associated membrane proteins (LAMP). However, roughly one-fourth of the bacteria taken up evade this initial killing event, remaining in nonacidic compartments. Forty-eight hours after infection, the number of intracellular bacteria per cell increased, suggesting that B. pertussis is capable of replicating in this type of compartment. Viable bacteria accumulated within phagosomal compartments positive for the early endosomal marker Rab5 but not the late endosomal marker LAMP. Moreover, B. pertussis-containing phagosomes acquired exogenously added transferrin, indicating that intracellular bacteria have access to extracellular components and essential nutrients via the host cell recycling pathway. Overall, these results suggest that B. pertussis survives and eventually replicates in compartments with characteristics of early endosomes, potentially contributing to its extraordinary ability to persist within hosts and populations.

scholarly journals Endocytic Host Cell Machinery Plays a Dominant Role in Intracellular Trafficking of Incoming Human Immunodeficiency Virus Type 1 in Human Placental Trophoblasts

2004 ◽  
Vol 78 (21) ◽  
pp. 11904-11915 ◽  
Author(s):  
Gaël Vidricaire ◽  
Michael Imbeault ◽  
Michel J. Tremblay
Keyword(s):  
Human Immunodeficiency Virus ◽  
Host Cell ◽  
Human Immunodeficiency Virus Type ◽  
Virus Type ◽  
Intracellular Trafficking ◽  
Confocal Imaging ◽  
Early Endosomes ◽  
Immunodeficiency Virus ◽  
Hiv 1

ABSTRACT Vertical transmission of human immunodeficiency virus type 1 (HIV-1) is the primary cause of infection by this retrovirus in infants. In this study, we report for the first time that there is a correlation between endocytic uptake of HIV-1 and virus gene expression in polarized trophoblasts. To shed light on the relationship between endocytosis and the fate of HIV-1 in polarized trophoblasts, the step-by-step movements of HIV-1 within the endocytic compartments were tracked by confocal imaging. Incoming virions were initially located in early endosomes. As time progressed, virions accumulated in late endosomes. HIV-1 was also found in apical recycling endosomes and at the basolateral pole. Experiments performed with indicator cells revealed that HIV-1 is recycled and transcytosed. These data indicate that the intracellular trafficking of HIV-1 upon entry into polarized human trophoblasts is a complex process which requires the active participation of the endocytic host cell machinery.

scholarly journals Chikungunya virus requires an intact microtubule network for efficient viral genome delivery

2020 ◽  
Vol 14 (8) ◽  
pp. e0008469
Author(s):  
Tabitha E. Hoornweg ◽  
Ellen M. Bouma ◽  
Denise P.I. van de Pol ◽  
Izabela A. Rodenhuis-Zybert ◽  
Jolanda M. Smit
Keyword(s):  
Viral Genome ◽  
Chikungunya Virus ◽  
Microtubule Network

scholarly journals How Influenza Virus Uses Host Cell Pathways during Uncoating

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1722
Author(s):  
Etori Aguiar Moreira ◽  
Yohei Yamauchi ◽  
Patrick Matthias
Keyword(s):  
Host Cell ◽  
Conformational Changes ◽  
Viral Genome ◽  
Human Population ◽  
Influenza A ◽  
Virus Genome ◽  
Cell Entry ◽  
Infection Cycle ◽  
Late Endosomes ◽  
Host Cell Entry

Influenza is a zoonotic respiratory disease of major public health interest due to its pandemic potential, and a threat to animals and the human population. The influenza A virus genome consists of eight single-stranded RNA segments sequestered within a protein capsid and a lipid bilayer envelope. During host cell entry, cellular cues contribute to viral conformational changes that promote critical events such as fusion with late endosomes, capsid uncoating and viral genome release into the cytosol. In this focused review, we concisely describe the virus infection cycle and highlight the recent findings of host cell pathways and cytosolic proteins that assist influenza uncoating during host cell entry.

An Integrated Chikungunya Virus Database to Facilitate Therapeutic Analysis: ChkVDb

2019 ◽  
Vol 14 (4) ◽  
pp. 323-332 ◽  
Author(s):  
Priya Narang ◽  
Mehak Dangi ◽  
Deepak Sharma ◽  
Alka Khichi ◽  
Anil Kumar Chhillar
Keyword(s):  
Side Effects ◽  
Chikungunya Virus ◽  
Cpg Island ◽  
Antiviral Treatment ◽  
Current Treatment ◽  
T Cell Epitopes ◽  
Entire Genome ◽  
Viral Pathogen ◽  
Related Data ◽  
Codon Context

Background: Chikungunya infection flare-ups have manifested in nations of Africa, Asia, and Europe including Indian and Pacific seas. It causes fever and different side effects include muscle torment, migraine, sickness, exhaustion and rash. It has turned into another, startling general medical issue in numerous tropical African and Asian countries and is presently being viewed as a genuine risk. No antiviral treatment or vaccine is yet available for this ailment. The current treatment is centered just on mitigating its side effects. Objective: The objective was to encourage the study on this viral pathogen, by the development of a database dedicated to Chikungunya Virus, that annotates and unifies the related data from various resources. associations while known disease-lncRNA associations are required only. Method: It undertook a consolidated approach for Chikungunya Virus genomic, proteomic, phylogenetics and therapeutic learning, involving the entire genome sequences and their annotation utilizing different in silico tools. Annotation included the information for CpG Island, usage bias, codon context and phylogenetic analysis at both the genome and proteome levels. Results: This database incorporates information of 41 strains of virus causing Chikungunya infection that can be accessed conveniently as well as downloaded effortlessly. Therapeutics section of this database contains data about B and T cell Epitopes, siRNAs and miRNAs that can be used as potential therapeutic targets. Conclusion: This database can be explored by specialists and established researchers around the world to assist their research on this non-treatable virus. It is a public database available from “www.chkv.in”.</P>

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