Formation of DNA replication structures in herpes virus-infected cells requires a viral DNA binding protein

Kops, Anne de Bruyn; Knipe, David M.

doi:10.1016/0092-8674(88)90141-9

Cited by 243 publications

(66 citation statements)

References 50 publications

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“…In this case, the viral capsids were trapped in the nucleus, indicating the blockade of nuclear maturation, similar to that described for the s309 mutant (43). Furthermore, the nuclear capsids in control cells were dispersed and surrounded an intranuclear structure of altered electron density reminiscent of replication compartments (25) (outlined in Fig. 3A and C).…”

Section: Resultsmentioning

confidence: 61%

Characterization of Conserved Region 2-Deficient Mutants of the Cytomegalovirus Egress Protein pM53

Pogoda

Bosse

Wagner

et al. 2012

J Virol

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; therefore, the present study describes the construction of M53 alleles lacking CR2 (either completely or partially) and subsequent examination of the DN effect on MCMV replication upon conditional expression. Overexpression of CR2-deficient pM53 inhibited virus production by about 10,000-fold. This was due to interference with capsid export from the nucleus and viral genome cleavage/packaging. In addition, the fate of the nuclear envelopment complex in the presence of DN pM53 overexpression was analyzed. The CR2 mutants were able to bind to pM50, albeit to a lesser extent than the wild-type protein, and relocalized the wild-type nuclear envelope complex in infected cells. Unlike the CR4 DN, the CR2 DN mutants did not affect the stability of pM50.T he production and release of infectious herpesviral particles is a multistep process that begins in the nucleus of the infected cell, where viral genomes are packaged into nucleocapsids. A recent study of pseudorabies virus (PrV) showed that the nuclear envelope breaks down to allow the capsid to exit the nucleus (23). However, many studies have shown that the widely accepted nuclear egress pathway requires capsid translocation through the nuclear membrane via primary envelopment and de-envelopment. This process is facilitated by a number of viral and host cell protein interactions. From the viral side, the two major proteins involved are the gene products of UL31 and UL34 of HSV-1 (pUL31 and pUL34, respectively) or their homologues in all other herpesviruses studied to date, which promote efficient primary capsid envelopment at the inner nuclear membrane (INM) (11,24,40,50). The pUL31 homologue of murine cytomegalovirus (MCMV), pM53, is distributed throughout the nucleosol in the absence of other viral proteins. pM53 is targeted to the INM after interacting with the membrane protein pM50 (the homologue of pUL34) via a mechanism that is conserved throughout herpesvirus morphogenesis (13,27,31,46,55,56,62). pM50 and pM53 form the nuclear envelopment complex (NEC) and recruit other cellular and viral proteins, such as protein kinase C and pUS3 (3,8,22,37,40,41,47,53), resulting in displacement of the rigid nuclear lamina and nucleocapsid budding (reviewed in references 19 and 36).The mechanism by which the pUL31 and pUL34 homologues mediate capsid transition through the nuclear envelope is still not fully understood, but disruption of either partner usually leads to retention of viral capsids within the nucleus (5,11,13,24,31,50,64). Insights into this mechanism were gained from studying the PrV proteins pUL31 and pUL34, which induce vesicle formation in the nuclei of transfected cells in the absence of other viral components (20), supporting their role in the capsid envelopment process at the INM. The reciprocal binding sites, which result in NEC targeting to the INM, are well characterized in pM53 and pM50 and are conserved throughout the pUL31 family (5,13,14,30,31,54,56). Furthermore, recent studies of HSV-1 suggest a second essential interaction between the N-term...

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Section: Resultsmentioning

confidence: 61%

Characterization of Conserved Region 2-Deficient Mutants of the Cytomegalovirus Egress Protein pM53

Pogoda

Bosse

Wagner

et al. 2012

J Virol

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“…The cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal calf serum (FCS) and penicillin (100 IU/ml)-streptomycin (100 μg/ml) and amphotericin B (2.5 μg/ml) (de Bruyn and Knipe, 1988). ZIKV strains MR766 (Kuno and Chang, 2007) was obtained from ATCC.…”

Section: Methodsmentioning

confidence: 99%

Molecular cloning and characterization of the genes encoding the proteins of Zika virus

Hou

Cruz-Cosme

Armstrong

et al. 2017

Gene

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Zika virus (ZIKV) encodes a precursor protein (also called polyprotein) of about 3424 amino acids that is processed by proteases to generate 10 mature proteins and a small peptide. In the present study, we characterized the chemical features, suborganelle distribution and potential function of each protein using Flag-tagged protein expression system. Western blot analysis revealed the molecular weight of the proteins and the polymerization of E, NS1, and NS3 proteins. In addition, we performed multi-labeled fluorescent immunocytochemistry and subcellular fractionation to determine the subcellular localization of these proteins in host cells. We found that 1) the capsid protein colocalizes with 3 different cellular organelles: nucleoli, Golgi apparatus, and lipid droplet; NS2b and NS4a are associated with the Golgi apparatus; 2) the capsid and NS1proteins distribute in both cytoplasm and nucleus, NS5 is a nuclear protein; 3) NS3 protein colocalizes with tubulin and affects Lamin A; 4) Envelope, PrM, and NS2a proteins co-localize with the endoplasmic reticulum; 5) NS1 is associated with autophagosomes and NS4b is related to early endosome; 6) NS5 forms punctate structures in the nucleus that associate with splicing compartments shown by SC35, leading to reduction of SC35 protein level and trafficking of SC35 from the nucleus to the cytoplasm. These data suggest that ZIKV generates 10 functional viral proteins that exhibit distinctive subcellular distribution in host cells.

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“…However, it is conceivable that Py DNA replication also takes place in such a structure on the nuclear matrix, since the DNA replication of herpes simplex virus and adenovirus takes place in the subnuclear structure (33,53). If so, targeting of the replication origin to the replication factory would be an important step for the initiation of Py DNA replication.…”

Section: Discussionmentioning

confidence: 99%

The Capacity of Polyomavirus Enhancer Binding Protein 2αB (AML1/Cbfa2) To Stimulate Polyomavirus DNA Replication Is Related to Its Affinity for the Nuclear Matrix

Chen

Ito

Murakami

et al. 1998

Molecular and Cellular Biology

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The nuclear matrix is thought to play an important role in the DNA replication of eukaryotic cells, although direct evidence for such a role is still lacking. A nuclear matrix-associated transcription factor, polyomavirus (Py) enhancer binding protein 2␣B1 (PEBP2␣B1) (AML1/Cbfa2), was found to stimulate Py replication through its cognate binding site. The minimal replication activation domain (RAD) was identified between amino acid (aa) 302 and aa 371 by using a fusion protein containing the GAL4 DNA binding domain (GAL4-RAD). In addition, the region showed affinity for the nuclear matrix and, on the basis of competition studies, binding activity for one or more proteins involved in the initiation of Py DNA replication. A leukemogenic chimeric protein, AML1/ETO(MTG8), which does not contain this region of PEBP2␣B1/AML1, was also localized in the nuclear matrix fraction and competed for nuclear matrix association with PEBP2␣B1 and GAL4-RAD. Moreover, AML1/ETO inhibited Py DNA replication stimulated by PEBP2␣B1 and GAL4-RAD. The inhibition was specific for replication mediated by PEBP2␣B1 and GAL4-RAD, and proportional to the degree of loss of these activators from the nuclear matrix, suggesting a requirement for nuclear matrix targeting in the stimulation of Py DNA replication by RAD. These results are the first to suggest a molecular link between the initiation of DNA replication and the nuclear matrix compartment.Accumulating evidence suggests an involvement of transcription factors in the regulation of DNA replication in eukaryotic cells. The polyomavirus (Py) DNA replication system is ideal for elucidating the roles of transcription factors in DNA replication, as Py DNA replicates in the nuclei of rodent cells and relies entirely on host factors, except for a single viral protein, large T antigen

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Formation of DNA replication structures in herpes virus-infected cells requires a viral DNA binding protein

Cited by 243 publications

References 50 publications

Characterization of Conserved Region 2-Deficient Mutants of the Cytomegalovirus Egress Protein pM53

Characterization of Conserved Region 2-Deficient Mutants of the Cytomegalovirus Egress Protein pM53

Molecular cloning and characterization of the genes encoding the proteins of Zika virus

The Capacity of Polyomavirus Enhancer Binding Protein 2αB (AML1/Cbfa2) To Stimulate Polyomavirus DNA Replication Is Related to Its Affinity for the Nuclear Matrix

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