Characterization of the single-stranded DNA-binding domain of the herpes simplex virus protein ICP8

Leinbach, Susan S.; Heath, Louise S.

doi:10.1016/0167-4781(89)90017-1

Cited by 11 publications

(9 citation statements)

References 19 publications

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“…Since we have shown that incubation of ICP8 with a 20-nucleotide-long oligonucleotide results in a mixed population of singly and doubly ligated complexes, it could also be that steric hindrance at the interface between two molecules attached to the DNA strand influences the sensitivity to trypsin. The ICP8⌬N mutant was totally insoluble, suggesting that the C-terminal and N-terminal regions of ICP8 have a structurally well-defined interface, in agreement with previous evidence (24,39,72).…”

Section: Discussionsupporting

confidence: 80%

“…Limited proteolytic analysis (72) coupled with the characterization of temperature-sensitive (22) and deletion (24,38,39) mutants suggests putative boundaries of the minimal binding domain between residues 300 and 849, while more recent evidence, based on ICP8 photoaffinity labeling with oligonucleotides, indicated a slightly different region, namely, between residues 386 and 902 (73). Both stretches encompass a zinc finger motif predicted between amino acids 499 and 512, where the single ICP8 zinc atom is most likely bound.…”

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confidence: 99%

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The 60-Residue C-Terminal Region of the Single-Stranded DNA Binding Protein of Herpes Simplex Virus Type 1 Is Required for Cooperative DNA Binding

Mapelli

Mühleisen²,

Persico³

et al. 2000

J Virol

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Single-stranded (ssDNA) DNA binding proteins (SSBs) bind preferentially ssDNA in stoichiometric quantities with respect to their substrate, displaying little sequence preference and no associated ATPase activity (11). The binding is typically cooperative, though the level of cooperativity varies widely. Much effort has been spent on elucidation of the structural mechanism accounting for the cooperativity and its functional implications in the case of the filamentous phage GVP (6, 68), the phage T4 gp32 (10, 33), the adenovirus DNA binding protein (DBP) (34), the Escherichia coli SSB (19,42), and the eukaryotic replication protein A (30, 31), while little is known about the SSBs of the Herpesviridae.The herpes simplex virus type 1 (HSV-1) SSB, infected cell polypeptide 8 (ICP8), is a 128-kDa nuclear zinc metalloprotein encoded by the UL29 gene (22). By virtue of its ability to bind oligonucleotide as well as to mediate specific protein-protein interactions, it was shown to have essential functions in DNA metabolism during the viral lytic cycle. ICP8 is one of the seven ␤, or delayed-early, genes required for viral genome replication, which proceeds via a rolling circle mechanism (63, 65). Replication occurs in globular nuclear domains termed replication compartments (55) whose location is defined by the preexisting host cell nuclear architecture, most probably at the periphery of the nuclear matrix-associated ND10 domains, where the viral transactivator ICP0 and the viral input genome migrate in the early stages of infection (43, 47). Evidence has been provided that the seven essential proteins, the origin binding protein (OBP), the polymerase with its processivity factor (UL30 and UL42), the trimeric primase-helicase complex (UL52, UL5, and UL8), and ICP8, accumulate in punctuate prereplicative sites for the assembly of the multiprotein complex, or primosome, which promotes efficient genomic replication (40,71,76). During initiation, ICP8 associates with the carboxy-terminal domain of the OBP at the origin of replication to assist the ATP-dependent bidirectional origin unwinding (3,36,37,45). It enhances the polymerase processivity (29, 50, 61) and stimulates the helicase-primase activity via interaction with the UL8 subunit (4,17,21). In accord with its ability to destabilize DNA helices (5) and promote Mg-dependent complementary-strand renaturation (16), ICP8 can catalyze homologous pairing and strand transfer (7), and hence it participates in the frequent DNA recombination events. Genetic evidence implies a role for ICP8 in the regulation of viral gene expression (12,26), in agreement with the reported ICP8 affinity for polyriboadenylate (61).The DNA binding properties of ICP8 have been extensively investigated. ICP8 binds ssDNA rapidly and cooperatively, with a modest preference for the HSV genomic GC-rich sequences, holding the nucleotide filaments in an extended conformation (35,58). Optimal binding occurs at neutral pH and 150 mM salt concentration (61). Based on filter binding assays (58), nuclease prote...

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Section: Discussionsupporting

confidence: 80%

mentioning

confidence: 99%

The 60-Residue C-Terminal Region of the Single-Stranded DNA Binding Protein of Herpes Simplex Virus Type 1 Is Required for Cooperative DNA Binding

Mapelli

Mühleisen²,

Persico³

et al. 2000

J Virol

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“…Gao et al analysed a variety of ICP 8 mutants and reported that the ss DNA-binding domain was contained in the C-terminal half of the ICP 8 (residues 564-1081), while the N-terminus was also required for tight and efficient binding [12]. Similar results were also obtained by Leinbach and Heath [23,24]. Gao et al also showed that a region of residues 348-450 might be involved in binding to DNA and that numerous [3-strands predicted for this region could fold to form a channel for binding to ss DNA [111.…”

Section: Predicted Amino Acid Sequence Of the Hsv-2 Icp 8 And Comparisupporting

confidence: 76%

Nucleotide sequence of the major DNA-binding protein gene of herpes simplex virus type 2 and a comparison with the type 1 counterpart

Toh

Liu

Tanaka

et al. 1993

Archives of Virology

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The nucleotide sequence of a region encompassing about 5,200 base pairs (bp) of the left side of the origin of replication in the long unique region of the herpes simplex virus type 2 (HSV-2) has been determined. This region contained the major DNA-binding protein or the infected-cell protein 8 (ICP 8) gene and 5'-part of the counterpart of HSV-1 ICP 18.5 gene. A comparison of the nucleotide sequence of the ICP8 gene between HSV-1 and HSV-2 showed an 89.8% homology. A primer extension analysis for the HSV-2 ICP 8 mRNA showed that the major transcriptional start site was mapped at 315 bp upstream of the initiation codon. A comparison of the predicted functional amino acid sequence of the ICP 8 between HSV-1 and HSV-2 revealed a striking homology (97.2%), the value of which was the highest among those of the other polypeptides encoded by HSV-1 and HSV-2. Some domains, which were shown to be required for the nuclear function, the binding to single-stranded DNA and the nuclear localization were well conserved. In addition, the nucleotide and the functional amino acid sequences of a part of the HSV-2 counterpart of the HSV-1 ICP 18.5 gene were also compared, demonstrating an 88.4% and 95.9% homology, respectively.

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“…We identified a 56-kDa peptide which binds DNA and exhibits the expected specificity for single-stranded DNA. This fragment overlaps two sites within the central portion of the molecule which have recently been shown by others to bind DNA (17,33). Within this peptide, we identified certain regions which may participate directly in DNA binding based on sequence comparisons with other DNA-binding proteins.…”

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confidence: 96%

Characterization of a major DNA-binding domain in the herpes simplex virus type 1 DNA-binding protein (ICP8)

Wang

Hall

1990

J Virol

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We have studied the major DNA-binding protein (ICP8) from herpes simplex virus type 1 to identify its DNA-binding site. Since we obtained our protein from a cell line carrying multiple chromosomally located copies of the ICP8 gene, we first analyzed this protein to assess its similarity to the corresponding viral protein. Our protein resembled the viral protein by molecular weight, response to antibody, preference for binding single-stranded DNA, and ability to lower the melting temperature of poly(dA-dT). To define the DNA-binding domain, we subjected the protein to limited trypsin digestion and separated the peptide products on a sodium dodecyl sulfate-polyacrylamide gel. These fragments were then transferred to a nitrocellulose membrane, renatured in situ, and tested for their ability to bind DNA. From this assay, we identified four fragments which both bound DNA and exhibited the expected binding preference for single-stranded DNA. The sequence of the smallest of these fragments was determined and corresponds to a polypeptide spanning residues 300 to 849 in the intact protein. This peptide contains several regions which may be important for DNA binding based on sequence similarities in single-stranded DNA-binding proteins from other herpesviruses and, in one case, on a conserved sequence found in more distant procaryotic and eucaryotic proteins.

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Characterization of the single-stranded DNA-binding domain of the herpes simplex virus protein ICP8

Cited by 11 publications

References 19 publications

The 60-Residue C-Terminal Region of the Single-Stranded DNA Binding Protein of Herpes Simplex Virus Type 1 Is Required for Cooperative DNA Binding

The 60-Residue C-Terminal Region of the Single-Stranded DNA Binding Protein of Herpes Simplex Virus Type 1 Is Required for Cooperative DNA Binding

Nucleotide sequence of the major DNA-binding protein gene of herpes simplex virus type 2 and a comparison with the type 1 counterpart

Characterization of a major DNA-binding domain in the herpes simplex virus type 1 DNA-binding protein (ICP8)

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