Monoclonal Antibodies Against Herpes Simplex Virus Type 1-infected Nuclei Defining and Localizing the ICP8 Protein, 65K DNA-binding Protein and Polypeptides of the ICP35 Family

Schenck, P.; Pietschmann, S.; Gelderblom, H.; Pauli, Georg; Ludwig, Hans

doi:10.1099/0022-1317-69-1-99

Cited by 26 publications

(18 citation statements)

References 13 publications

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“…Aliquots of lysates prepared from 10& cells were incubated for 18 h at 4 mC with one of the following : rabbit polyclonal anti-ICP27 antibody, rabbit polyclonal anti-US11 antibody (Diaz et al, 1993), mouse monoclonal anti-DBP antibody or mouse CHGG monoclonal antibody directed against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Interchim), each diluted 50-fold in lysis buffer. Anti-ICP27 and anti-DBP antibodies (antibody 42 and antibody Z1F11 directed against the DBP of 65 kDa, respectively) were kindly provided by H. Marsden (MRC, Glasgow, UK) (Schenck et al, 1988 ;Sinclair et al, 1994). Antigen-antibody complexes were collected with protein A coupled to Sepharose CL-4B beads (Pharmacia) for 30 min at 4 mC.…”

Section: Methodsmentioning

confidence: 99%

Translational control of viral and host protein synthesis during the course of herpes simplex virus type 1 infection: evidence that initiation of translation is the limiting step.

Laurent¹,

Madjar²,

Greco³

1998

Journal of General Virology

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Herpes simplex virus type 1 (HSV-1) infection induces the selective shut-off of host protein synthesis, other than ribosomal proteins, and the successive synthesis of viral proteins. Because viral mRNAs persist in the cytoplasm after viral protein synthesis has been inhibited, we hypothesized that viral gene expression may be under translational control. Expression of genes encoding immediate early ICP27, early DBP and late US11 proteins, together with glyceraldehyde-3-phosphate dehydrogenase (GAPDH), was monitored over the course of infection at the level of mRNA and protein synthesis. After an efficient synthesis beginning with the appearance of successive viral mRNAs in the cytoplasm, synthesis of viral proteins was shut off similarly to the synthesis of GAPDH. This shut-off

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Section: Methodsmentioning

confidence: 99%

Translational control of viral and host protein synthesis during the course of herpes simplex virus type 1 infection: evidence that initiation of translation is the limiting step.

Laurent¹,

Madjar²,

Greco³

1998

Journal of General Virology

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“…The membranes were incubated with a 500ϫ dilution of either a rabbit polyclonal antinucleolin antibody (19); antifibrillarin (N-15; Santa Cruz), anti-ICP27, or anti-US11 antibodies (14); or mouse monoclonal anti-B23 (H-106; Sigma), anti-␤-actin (AC-15; Sigma), antihistone H3 (Abcam), or anti-UL42 antibodies. Anti-ICP27 and anti-UL42 antibodies were kindly provided by H. Marsden (antibodies 42 and Z1F11, respectively) (53,58). Proteins were revealed by chemiluminescence (ECL from Amersham Biosciences) using an anti-rabbit or an anti-mouse peroxidase-conjugated antibody (Sigma) or an anti-goat peroxidase-conjugated antibody (Santa Cruz), diluted 1:10,000.…”

Section: Methodsmentioning

confidence: 99%

Nucleolin Is Required for an Efficient Herpes Simplex Virus Type 1 Infection

Callé

Ugrinova

Epstein

et al. 2008

J Virol

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Productive infection by herpes simplex virus type 1 (HSV-1), which occurs in the host cell nucleus, is accompanied by dramatic modifications of the nuclear architecture, including profound alterations of nucleolar morphology. Here, we show that the three most abundant nucleolar proteins-nucleolin, B23, and fibrillarin-are redistributed out of the nucleoli as a consequence of HSV-1 infection. We show that the amount of nucleolin increases progressively during the course of infection. We demonstrate for the first time that a nucleolar protein, i.e., nucleolin, colocalizes with ICP8 in the viral replication compartments, at the time when viral replication is effective, suggesting an involvement of nucleolin in the HSV-1 DNA replication process. At later times of infection, a granular form of nucleolin localizes to the cytoplasm, in structures that display the characteristic features of aggresomes, indicating that this form of nucleolin is very probably destined for degradation. The delocalization of nucleolin from the nucleoli requires the viral ICP4 protein or a factor(s) whose expression involves ICP4. Using small interfering RNA technology, we show that viral replication requires a high level of nucleolin expression, demonstrating for the first time a direct role for a nucleolar protein in herpes simplex virus biology.Herpes simplex virus type 1 (HSV-1) is a human herpesvirus consisting of an outer envelope, a tegument, a capsid, and a linear double-stranded DNA. Productive infection consists of a highly ordered program of viral gene expression, DNA replication, and virion assembly that leads to the formation of infectious viral progeny and cell death (48). The viral DNA contains at least 80 genes whose expression is sequentially and temporally regulated by complex regulatory mechanisms. Viral genes can be divided into immediate-early, early, and late genes, according to their kinetics of expression. Proteins encoded by immediate-early genes are involved in the regulation of the synthesis of early and late proteins. Proteins encoded by early genes participate in viral DNA replication, and proteins encoded by late genes are mainly the structural components of the viral particles.Viral transcription, DNA replication, assembly of new capsids, and packaging of HSV-1 DNA occur in the host cell nucleus. As a consequence, HSV-1-infected cells undergo a variety of changes including dramatic modifications of the nuclear architecture. The formation of viral replication compartments (VRC), which are the sites of replication, transcription, and encapsidation of HSV-1 genomes, is accompanied by the marginalization of chromatin and the disruption of the nuclear lamina and of PML bodies, as well as by a profound modification of nucleolar morphology (2,15,33,39). Soon after infection, nucleoli increase in size, localize close to the nuclear membrane, and finally become fragmented in small pieces (39,49). In addition, several viral proteins, including ICP0, ICP4, ICP27, US11, and gamma 34.5, localize at least transiently to nucl...

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“…The physical interaction assay was performed in microtitre wells coated with pol using MAb ZIFII !24~ (Schenk et al, 1988) to detect UL42 protein binding as described …”

Section: Methodsmentioning

confidence: 99%

The Herpes Simplex Virus Type 1 UL8 Protein Influences the Intracellular Localization of the UL52 but not the ICP8 or POL Replication Proteins in Virus-infected Cells

Marsden¹,

Cross²,

Francis³

et al. 1996

Journal of General Virology

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We have developed a panel of 14 monoclonal antibodies (MAbs) to POL, the catalytic subunit of herpes simplex virus type 1 (HSV-1) DNA polymerase encoded by gene UL30, and one HAb to the UL52 protein, another of the seven proteins essential for replication of HSV DNA. The approximate locations of the epitopes of the polymerase-specific MAbs were identified using truncated polymerase molecules, and the antibodies were characterized in a number of immunological assays allowing eight different specificities to be recognized. These MAbs, together with a polyclonal antibody raised in rabbits against a third DNA replication protein, ICP8, were used to localize the respective proteins by immunofluorescence in cells infected with wild-type HSV-1 or the DNA replication-defective mutants ombUL8 or 2-2. In BHK cells infected with ambULS, a mutant with an amber termination codon within the coding region of gene UL8, the UL52 protein did not enter the nucleus, although ICP8 and POL entered the nucleus in a normal fashion. The failure of the UL52 protein to be correctly transported to the nucleus was also observed in both HFL and Vero cells infected with ambUL8. In contrast, UL52 protein was transported to the nucleus in BHK cells infected with wild-type HSV-1 or with 2-2, a mutant lacking a functional UL9 protein.

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Monoclonal Antibodies Against Herpes Simplex Virus Type 1-infected Nuclei Defining and Localizing the ICP8 Protein, 65K DNA-binding Protein and Polypeptides of the ICP35 Family

Cited by 26 publications

References 13 publications

Translational control of viral and host protein synthesis during the course of herpes simplex virus type 1 infection: evidence that initiation of translation is the limiting step.

Translational control of viral and host protein synthesis during the course of herpes simplex virus type 1 infection: evidence that initiation of translation is the limiting step.

Nucleolin Is Required for an Efficient Herpes Simplex Virus Type 1 Infection

The Herpes Simplex Virus Type 1 UL8 Protein Influences the Intracellular Localization of the UL52 but not the ICP8 or POL Replication Proteins in Virus-infected Cells

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