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Egress of HSV-1 capsid requires the interaction of VP26 and a cellular tetraspanin membrane protein.

Wang L, Liu L, Che Y, Wang L, Jiang L, Dong C, Zhang Y, Li Q - Virol. J. (2010)

Bottom Line: In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro.The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication.Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute Of Medical Biology, Chinese Academy of Medicine Science, Peking Union Medical College, Kunming 650118, P. R. China.

ABSTRACT
HSV-1 viral capsid maturation and egress from the nucleus constitutes a self-controlled process of interactions between host cytoplasmic membrane proteins and viral capsid proteins. In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro. The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication. Additionally, analysis of a recombinant virus HSV-1-UG showed that mutating VP26 resulted in a decreased viral replication rate and in aggregation of viral mutant capsids in the nucleus. Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

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Intracellular analysis of CTMP-7 in HSV-1 infected cells. Vero cells were transiently transfected with plasmid pGFP-CTMP or pGFP-N followed by HSV-1 infection at 1 MOI, and then observed at 24 h post transfection by fluorescence microscopy without fixation. At 30 h post-infection, the cells were trypsinized, washed in PBS and analyzed by flow cytometry. a. Observation of Vero cells transfected with pGFP-CTMP or pGFP-N and infected by HSV1 at 24 h post-infection under fluorescence microscope. Upper row: Cells transfected with pGFP-CTMP and infected by HSV-I. Lower row: Cells transfected with pGFP-N and infected by HSV-I. b. Cytometric analysis of Vero cells transfected with pGFP-CTMP and pGFP-N followed by HSV-1 infection or not at 30 h post-infection. Column 1: Cells transfected with pGFP-N, upper - cells HSV-I infection free; lower - cells at 30 h post-infection. Column 2: Control cells transfected with pGFP-CTMP with HSV1 infection free, upper is cell at the same time point to 16 h post-infection of infective example, lower is cells at the same time point to 30 h post-infection of infective example. Column 3: Cells transfected with pGFP-CTMP and followed by HSV1 infection, upper is cell at 16 h post-infection; down is cell at 30 h post-infection.
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Figure 2: Intracellular analysis of CTMP-7 in HSV-1 infected cells. Vero cells were transiently transfected with plasmid pGFP-CTMP or pGFP-N followed by HSV-1 infection at 1 MOI, and then observed at 24 h post transfection by fluorescence microscopy without fixation. At 30 h post-infection, the cells were trypsinized, washed in PBS and analyzed by flow cytometry. a. Observation of Vero cells transfected with pGFP-CTMP or pGFP-N and infected by HSV1 at 24 h post-infection under fluorescence microscope. Upper row: Cells transfected with pGFP-CTMP and infected by HSV-I. Lower row: Cells transfected with pGFP-N and infected by HSV-I. b. Cytometric analysis of Vero cells transfected with pGFP-CTMP and pGFP-N followed by HSV-1 infection or not at 30 h post-infection. Column 1: Cells transfected with pGFP-N, upper - cells HSV-I infection free; lower - cells at 30 h post-infection. Column 2: Control cells transfected with pGFP-CTMP with HSV1 infection free, upper is cell at the same time point to 16 h post-infection of infective example, lower is cells at the same time point to 30 h post-infection of infective example. Column 3: Cells transfected with pGFP-CTMP and followed by HSV1 infection, upper is cell at 16 h post-infection; down is cell at 30 h post-infection.

Mentions: The fusion protein of CTMP-7 and GFP encoded by eukaryotic expression vector pGFP-CTMP was expressed by transient transfection in Vero cells. Visualization of the expressed CTMP-7-GFP fusion protein in normal cells revealed aggregation in punctate spots on the nuclear and cytoplasmic membranes similar to the pattern exhibited by other tetraspanin family members (Fig.2a, left at upper)[21]. In contrast, the fluorescent punctate distribution was altered drastically in HSV-1 infected Vero cells expressing the CTMP-7-GFP fusion protein. Bright punctate spots were distributed from the nuclear membrane to the cytoplasm at 24 h post-infection (Fig.2a, right at upper). Compared with control cells expressing fluorescent GFP protein where no significant alteration was visualized (Fig 2a down row). Flow cytometric analysis showed an obvious decrease of fluorescence intensity in cells expressing tetraspanin-GFP fusion protein at 30 h post HSV-1 infection (Fig.2b).


Egress of HSV-1 capsid requires the interaction of VP26 and a cellular tetraspanin membrane protein.

Wang L, Liu L, Che Y, Wang L, Jiang L, Dong C, Zhang Y, Li Q - Virol. J. (2010)

Intracellular analysis of CTMP-7 in HSV-1 infected cells. Vero cells were transiently transfected with plasmid pGFP-CTMP or pGFP-N followed by HSV-1 infection at 1 MOI, and then observed at 24 h post transfection by fluorescence microscopy without fixation. At 30 h post-infection, the cells were trypsinized, washed in PBS and analyzed by flow cytometry. a. Observation of Vero cells transfected with pGFP-CTMP or pGFP-N and infected by HSV1 at 24 h post-infection under fluorescence microscope. Upper row: Cells transfected with pGFP-CTMP and infected by HSV-I. Lower row: Cells transfected with pGFP-N and infected by HSV-I. b. Cytometric analysis of Vero cells transfected with pGFP-CTMP and pGFP-N followed by HSV-1 infection or not at 30 h post-infection. Column 1: Cells transfected with pGFP-N, upper - cells HSV-I infection free; lower - cells at 30 h post-infection. Column 2: Control cells transfected with pGFP-CTMP with HSV1 infection free, upper is cell at the same time point to 16 h post-infection of infective example, lower is cells at the same time point to 30 h post-infection of infective example. Column 3: Cells transfected with pGFP-CTMP and followed by HSV1 infection, upper is cell at 16 h post-infection; down is cell at 30 h post-infection.
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Related In: Results  -  Collection

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Figure 2: Intracellular analysis of CTMP-7 in HSV-1 infected cells. Vero cells were transiently transfected with plasmid pGFP-CTMP or pGFP-N followed by HSV-1 infection at 1 MOI, and then observed at 24 h post transfection by fluorescence microscopy without fixation. At 30 h post-infection, the cells were trypsinized, washed in PBS and analyzed by flow cytometry. a. Observation of Vero cells transfected with pGFP-CTMP or pGFP-N and infected by HSV1 at 24 h post-infection under fluorescence microscope. Upper row: Cells transfected with pGFP-CTMP and infected by HSV-I. Lower row: Cells transfected with pGFP-N and infected by HSV-I. b. Cytometric analysis of Vero cells transfected with pGFP-CTMP and pGFP-N followed by HSV-1 infection or not at 30 h post-infection. Column 1: Cells transfected with pGFP-N, upper - cells HSV-I infection free; lower - cells at 30 h post-infection. Column 2: Control cells transfected with pGFP-CTMP with HSV1 infection free, upper is cell at the same time point to 16 h post-infection of infective example, lower is cells at the same time point to 30 h post-infection of infective example. Column 3: Cells transfected with pGFP-CTMP and followed by HSV1 infection, upper is cell at 16 h post-infection; down is cell at 30 h post-infection.
Mentions: The fusion protein of CTMP-7 and GFP encoded by eukaryotic expression vector pGFP-CTMP was expressed by transient transfection in Vero cells. Visualization of the expressed CTMP-7-GFP fusion protein in normal cells revealed aggregation in punctate spots on the nuclear and cytoplasmic membranes similar to the pattern exhibited by other tetraspanin family members (Fig.2a, left at upper)[21]. In contrast, the fluorescent punctate distribution was altered drastically in HSV-1 infected Vero cells expressing the CTMP-7-GFP fusion protein. Bright punctate spots were distributed from the nuclear membrane to the cytoplasm at 24 h post-infection (Fig.2a, right at upper). Compared with control cells expressing fluorescent GFP protein where no significant alteration was visualized (Fig 2a down row). Flow cytometric analysis showed an obvious decrease of fluorescence intensity in cells expressing tetraspanin-GFP fusion protein at 30 h post HSV-1 infection (Fig.2b).

Bottom Line: In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro.The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication.Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute Of Medical Biology, Chinese Academy of Medicine Science, Peking Union Medical College, Kunming 650118, P. R. China.

ABSTRACT
HSV-1 viral capsid maturation and egress from the nucleus constitutes a self-controlled process of interactions between host cytoplasmic membrane proteins and viral capsid proteins. In this study, a member of the tetraspanin superfamily, CTMP-7, was shown to physically interact with HSV-1 protein VP26, and the VP26-CTMP-7 complex was detected both in vivo and in vitro. The interaction of VP26 with CTMP-7 plays an essential role in normal HSV-1 replication. Additionally, analysis of a recombinant virus HSV-1-UG showed that mutating VP26 resulted in a decreased viral replication rate and in aggregation of viral mutant capsids in the nucleus. Together, our data support the notion that biological events mediated by a VP26 - CTMP-7 interaction aid in viral capsid enveloping and egress from the cell during the HSV-1 infectious process.

Show MeSH
Related in: MedlinePlus