Visualization of the Annealing of Complementary Single-stranded DNA Catalyzed by the Herpes Simplex Virus Type 1 ICP8 SSB/Recombinase

Makhov, Alexander M.; Griffith, Jack D.

doi:10.1016/j.jmb.2005.11.022

Cited by 23 publications

(50 citation statements)

References 54 publications

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“…4E and F). Addition of up to a 10-fold higher concentration of UL8 (1 µM) failed to generate any regular extended nucleoprotein filaments of the kinds seen with ICP8 and Orf6 at high protein concentrations (14,36). The implications of the difference in DNA binding properties between UL8 and ICP8 will be explored in the Discussion.…”

Section: Resultsmentioning

confidence: 94%

“…Given the well-documented activity of ICP8 as an annealase (14,15,37), we tested UL8 for its ability to anneal complementary ssDNA. Linearized, blunt-ended 3987 bp pSAK dsDNA (38) (50 ng, 1 nM dsDNA, 10 14 nt) was heatdenatured to produce ssDNA, rapidly quenched on ice and then incubated in the presence or absence of UL8.…”

Section: Resultsmentioning

confidence: 99%

“…ICP8 forms helical protein filaments in the absence of DNA (12), and filament formation appears to be required for pre-replicative site and replication compartment formation (13). ICP8 also forms superhelical structures on ssDNA, and these structures are believed to promote annealing of complementary ssDNA (14,15). ICP8, in conjunction with the viral 5` to 3' exonuclease (UL12) has been shown to carry out a strand exchange reaction that is analogous to the activity of the lambda red alpha/beta proteins (16,17).…”

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confidence: 99%

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O-GlcNAc Transferase Regulates Mitotic Chromatin Dynamics

Sakabe

Hart

2010

Journal of Biological Chemistry

120

115

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During lytic infection, herpes simplex virus (HSV) DNA is replicated by a mechanism involving DNA recombination. For instance, replication of the HSV-1 genome produces X-and Y-branched structures, reminiscent of recombination intermediates. HSV-1`s replication machinery includes a trimeric helicase/primase is composed of helicase (UL5) and primase (UL52) subunits and a third subunit, UL8. UL8 has been reported to stimulate the helicase and primase activities of the complex in the presence of ICP8, an HSV-1 protein that functions as an annealase, a protein that binds complementary single-stranded (ss)DNA and facilitates its annealing to duplex DNA. UL8 also influences the intracellular localization of the UL5/UL52 subunits, but UL8`s catalytic activities are not known. In this study, we used a combination of biochemical techniques and transmission electron microscopy. First, we report that UL8 alone forms protein filaments in solution. Moreover, we also found that, UL8 binds to ssDNAs >50 nt long and promotes the annealing of complementary ssDNA to generate highly branched duplex DNA structures. Finally, UL8 has a very high affinity for replication fork structures containing a gap in the lagging strand as short as 15 nt, suggesting that UL8 may aid in directing or loading the trimeric complex onto a replication fork. The properties of UL8 uncovered here suggest that UL8 may be involved in the generation of the X-and Y-branched structures that are the hallmarks of HSV replication.Herpes simplex virus type 1 (HSV-1) has a 152-kb linear double-stranded (ds) DNA genome. During HSV-1 infection, the viral genome is released from the capsid into the nucleus and is replicated to form concatemeric DNA that is recognized by the packaging machinery to produce infectious virus. Seven virus-encoded proteins are required for viral DNA synthesis including an origin binding protein (UL9) and six core replication proteins: a two subunit DNA polymerase (UL30 and UL42), a three subunit helicase/primase complex (UL5, UL8 and UL52) and a multifunctional ssDNA binding protein (ICP8) (reviewed in ref.1). The six purified core replication proteins can function to produce linear concatemers in vitro if provided a primed template (2). On the other hand, in vivo, late replicating viral DNA has been reported to adopt a mixture of complex structures such as X-and Y-shaped branches and tangled masses, suggesting that recombination may play a role in the replication of HSV-1 DNA (3-7). Additional support for this concept was provided by Blümel et al. who showed that when SV40 genomes were replicated by the six HSV-encoded replication factors and SV40 large T antigen, concatemers composed of X-shaped DNA structures were observed (8). Since SV40 replication normally produces two circular daughter molecules, the complex DNA generated in the presence of HSV replication proteins suggests that the HSV replication machinery is inherently recombinogenic. A better understanding of the functions and properties of the essential viral replicatio...

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Section: Resultsmentioning

confidence: 94%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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O-GlcNAc Transferase Regulates Mitotic Chromatin Dynamics

Sakabe

Hart

2010

Journal of Biological Chemistry

120

115

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“…The process has been visualized demonstrating how two coiled ICP8-ssDNA nucleoprotein filaments generate intertwined coiled-coil structures in which strand annealing occurs (66). ICP8 has also been demonstrated to promote strand invasion in vitro, and together with helicaseprimase, it can promote strand exchange (67,68). A possible role could be to promote recombination-dependent DNA synthesis during repair of double-strand breaks (69,70).…”

Section: Properties Of Replisome Proteinsmentioning

confidence: 99%

Replication and Recombination of Herpes Simplex Virus DNA

Muylaert

Tang

Elias

2011

Journal of Biological Chemistry

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Replication of herpes simplex virus takes place in the cell nucleus and is carried out by a replisome composed of six viral proteins: the UL30-UL42 DNA polymerase, the UL5-UL8-UL52 helicase-primase, and the UL29 single-stranded DNA-binding protein ICP8. The replisome is loaded on origins of replication by the UL9 initiator origin-binding protein. Virus replication is intimately coupled to recombination and repair, often performed by cellular proteins. Here, we review new significant developments: the three-dimensional structures for the DNA polymerase, the polymerase accessory factor, and the singlestranded DNA-binding protein; the reconstitution of a functional replisome in vitro; the elucidation of the mechanism for activation of origins of DNA replication; the identification of cellular proteins actively involved in or responding to viral DNA replication; and the elucidation of requirements for formation of replication foci in the nucleus and effects on protein localization.Herpesviruses are found in all animals from molluscs to man. During evolution, the viruses have become tightly associated and co-evolved with their hosts. They seem to cross species borders only by accident, and in such rare instances, they may cause unexpected and severe disease. Herpesviruses have an unusual lifestyle; they cause lytic infection in cells, leading to efficient production of new infectious virus particles, and they also establish latent infections in either non-dividing neuronal cells or cycling cells of the immune system. The latent state is characterized by expression of a very limited set of genes to ascertain maintenance of virus chromosomes and to escape recognition by the immune system. The mechanisms for establishing latency appear to differ considerably for different herpesviruses. In contrast, mechanisms for replication of virus DNA during lytic infection and subsequent formation of infectious particles seem to be evolutionarily conserved. There is one notable exception; the mechanism for recognition of origins of DNA replication and initiation of DNA synthesis differs between the herpesvirus families.Humans can be infected by eight different herpesviruses. Herpes simplex viruses I and II and varicella zoster virus are alphaherpesviruses. Cytomegalovirus and the roseoloviruses, human herpesviruses 6 and 7, are classified as betaherpesviruses. Epstein-Barr virus and Kaposi's sarcoma-associated herpesvirus belong to the gammaherpesvirus subfamily.In this minireview, we discuss recent developments in replication, recombination, and repair of herpes simplex virus DNA. We will take as our starting point a previous minireview in the Journal of Biological Chemistry that provides an insightful and accurate description of basic mechanisms and components of the herpes simplex virus replication machinery (1). Noteworthy new developments have been (i) the presentation of three-dimensional structures for the DNA polymerase, the polymerase accessory factor, and the single-stranded DNA-binding protein (ssDNA) 2 ; (ii) the recons...

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“…Studies of RecT suggest that despite its independent evolution, it shares with RecA some of the hallmarks of a homologous pairing protein, including the ability to unstack ssDNA bases, which may facilitate homologous contacts (Noirot et al 2003). RecT shares the property with Redb and ICP8 of forming ringand filament-shaped structures in the presence of DNA (Thresher et al 1995;Passy et al 1999;Makhov and Griffith 2006;Makhov et al 2009). The ring form of Redb binds to ssDNA, whereas the helical form binds with higher affinity to the heteroduplex product of annealing (Karakousis et al 1998;Passy et al 1999), suggesting a mechanism for stabilizing the heteroduplex product of DNA strand exchange.…”

Section: Homologous Recombination By Exonucleases and Dna-annealing Pmentioning

confidence: 99%

DNA-Pairing and Annealing Processes in Homologous Recombination and Homology-Directed Repair

Morrical

2015

Cold Spring Harb Perspect Biol

114

127

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The formation of heteroduplex DNA is a central step in the exchange of DNA sequences via homologous recombination, and in the accurate repair of broken chromosomes via homology-directed repair pathways. In cells, heteroduplex DNA largely arises through the activities of recombination proteins that promote DNA-pairing and annealing reactions. Classes of proteins involved in pairing and annealing include RecA-family DNA-pairing proteins, single-stranded DNA (ssDNA)-binding proteins, recombination mediator proteins, annealing proteins, and nucleases. This review explores the properties of these pairing and annealing proteins, and highlights their roles in complex recombination processes including the double Holliday junction (DhJ) formation, synthesis-dependent strand annealing, and singlestrand annealing pathways-DNA transactions that are critical both for genome stability in individual organisms and for the evolution of species.

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Visualization of the Annealing of Complementary Single-stranded DNA Catalyzed by the Herpes Simplex Virus Type 1 ICP8 SSB/Recombinase

Cited by 23 publications

References 54 publications

O-GlcNAc Transferase Regulates Mitotic Chromatin Dynamics

O-GlcNAc Transferase Regulates Mitotic Chromatin Dynamics

Replication and Recombination of Herpes Simplex Virus DNA

DNA-Pairing and Annealing Processes in Homologous Recombination and Homology-Directed Repair

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