ssDNA-dependent colocalization of adeno-associated virus Rep and herpes simplex virus ICP8 in nuclear replication domains

Heilbronn, Regine; Engstler, Markus; Weger, Stefan; Krahn, Antje; Schetter, Christian; Boshart, Michael

doi:10.1093/nar/gkg827

Cited by 26 publications

(48 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To confirm the finding from the IP experiments, 3D deconvolution fluorescence microscopy with quantitative colocalization was carried out with WT1 and the HA-tagged hnRNP-U constructs (Heilbronn et al, 2003). Acquisition of the Z-axis stacks (0.2 mcm) of the images in either fluorescence channel was performed followed by Imaris-Colocalization quantitative analysis.…”

Section: Antibody (C Es Cells; D M15 Cells) (E)mentioning

confidence: 89%

hnRNP-U directly interacts with WT1 and modulates WT1 transcriptional activation

et al. 2006

View full text Add to dashboard Cite

The Wilms' tumour suppressor gene, WT1, encodes a zinc-finger protein that is mutated in Wilms' tumours and highly expressed in a wide variety of other malignancies. WT1 is a transcription factor that is likely to have additional, post-transcriptional, regulatory roles, although the molecular mechanisms by which WT1 acts remain poorly understood. We have combined genetic and biochemical approaches to show, that endogenous WT1 binds to heterogeneous nuclear ribonuclear protein U (hnRNP-U), that this interaction does not require any other proteins or nucleic acids, involves the zinc-fingers of WT1 and the middle domain of hnRNP-U, and that hnRNP-U can modulate WT1 transcriptional activation of a bona fide WT1 target gene. These findings increase our knowledge of how WT1 exerts its transcriptional regulatory role and suggests that hnRNP-U may be a candidate Wilms' tumour gene at 1q44.

show abstract

Section: Antibody (C Es Cells; D M15 Cells) (E)mentioning

confidence: 89%

hnRNP-U directly interacts with WT1 and modulates WT1 transcriptional activation

et al. 2006

View full text Add to dashboard Cite

show abstract

“…In analogy to the HSV-1 ori-binding protein UL9 [146], Rep probably triggers the assembly of the replication machinery by binding to the RBSs within the AAV-2 oris. In the next step, Rep68 and Rep78 are thought to recruit ICP8 through a direct protein-protein interaction, which in turn stimulates Rep DNA-binding and nicking activity [147,148]. The Rep68/78-ICP8 complex is then thought to recruit the helicase-primase complex, as well as the DNA polymerase holoenzyme.…”

Section: Hsv-1-based Gene Therapy Vectorsmentioning

confidence: 99%

“…Among the HSV-1 replication factors involved in AAV-2 replication, ICP8 can certainly be considered a key helper factor, since its presence is crucial to processive AAV-2 DNA replication. The function of ICP8 presumably lies in its ability to stabilize ssDNA and to hold the replication fork in an extended conformation [149], as well as in its ability to act as a scaffolding protein for the recruitment of the helicase-primase complex and the polymerase holoenzyme [90,147]. Consistent with a key role in the helper activity, the mere addition of purified ICP8 to extracts from uninfected HeLa cells is sufficient to support in vitro AAV-2 DNA replication [148].…”

Section: Hsv-1-based Gene Therapy Vectorsmentioning

confidence: 99%

“…Studying the interaction of replicating HSV-1 and AAV-2 DNA may therefore provide useful insight into the molecular mechanisms underlying the lower replication efficiency of HSV/AAV hybrid vectors compared with standard HSV-1 amplicon vectors. Analogous to the situation with other DNA viruses, AAV-2 DNA replication occurs in nuclear RCs, in which AAV-2 DNA co localizes with Rep proteins and components of the DNA Interactions between AAV-2 & HSV-1: implications for hybrid vector design Review replication machinery [28,147,148,151]. AAV-2 DNA replication initiates at small nuclear foci, so-called pre-replicative sites, which grow in size and appear to coalesce into large globular structures, eventually filling out most of the nucleus and displacing cellular chromatin to the nuclear periphery [14,28,151].…”

Section: Interactions At the Level Of Viral Replication Compartmentsmentioning

confidence: 99%

“…The HSV-1 IE gene products simultaneously transactivate HSV-1 early gene expression, resulting in the synthesis of a second set of helper factors, the ssDNA-binding protein ICP8, the helicase-primase complex and the DNA polymerase holoenzyme [138,144]. These helper factors are then thought to be recruited to the AAV-2 oris by a direct interaction of the large Rep proteins with ICP8 and to provide the replication machinery for AAV-2 DNA replication, the latter proceeding in RCs spatially separated from those of the helper virus [120,147,148,152]. The large Rep proteins also seem to have an intrinsic ability to inhibit the replication of cellular and HSV-1 DNA, which is dependent on their DNA-binding and ATPase/helicase activities [152,153,160,170].…”

Section: Conclusion and Future Perspective On The Analysis Of Aav-hsv Imentioning

confidence: 99%

See 2 more Smart Citations

Interactions Between AAV-2 and HSV-1: Implications for Hybrid Vector Design

Glauser

Fraefel

2011

Future Virol.

View full text Add to dashboard Cite

Herpes simplex virus type 1 (HSV-1)-based amplicon vectors have a transgene capacity of up to 150 kbp and can efficiently transduce many different cell types in culture and in vivo without causing cytopathic effects. However, these vectors do not support long-term transgene expression. Adeno-associated virus type 2 (AAV-2) has the capacity to integrate its genome into a specific site on human chromosome 19, but AAV-2-derived gene therapy vectors have a transgene capacity of only 4.5 kb. To combine the large transgene capacity of HSV-1 with the potential for site-specific genomic integration and long-term transgene expression of AAV-2, HSV/AAV hybrid vectors have been developed. This review describes the design, applications and limitations of these hybrid vectors. However, as HSV-1 is a full helper virus for AAV-2 replication, the main focus is the analysis of the molecular mechanisms of interaction between the two viruses. The knowledge of these interactions will have direct implications on the design of novel HSV/AAV hybrid vectors.

show abstract

Evidence for multiple signaling pathways in single squid olfactory receptor neurons

Mobley

Gandham

Lucero

2007

J of Comparative Neurology

View full text Add to dashboard Cite

At least two different G-protein-mediated transduction cascades, the adenylate cyclase and phospholipase C (PLC) pathway, process chemosensory stimuli for various species. In squid olfactory receptor neurons (ORNs), physiological studies indicate that both pathways may be present; however, confirmation of the transduction molecules at the protein level is absent. Here we provide evidence that the G-proteins involved in both adenylate cyclase and PLC pathways are present in squid ORNs (Lolliguncula brevis). We used immunoblotting to show that G␣ olf , G␣ q , and a downstream effector, enzyme PLC140, are present in the squid olfactory epithelium (OE). To localize these proteins to one or more of the five morphological cell types described for squid OE, paraformaldehyde-fixed olfactory organs were cryosectioned (10 m), double-labeled for G␣ olf , G␣ q , or PLC140, and imaged. Analysis of serial sections from entire olfactory organs for epithelial area and patterns of immunofluorescence revealed a region of highest immunoreactivity at the anterior half of the organ. At the cellular level, type 1 cells could not be distinguished morphologically and were not included in the analysis. The three labeling patterns observed in type 2 cells were G␣ q alone, PLC140 alone, and colocalization of G␣ q and PLC140. Subsets of cell types 3, 4, and 5 showed colocalization of G␣ olf with G␣ q but not with PLC140. These data suggest that the PLC pathway predominates in type 2 cells; however, coexpression of G␣ olf with G␣ q in cell types 3, 4, and 5 suggests that both pathways may participate in olfactory transduction in non-type 2 squid ORNs. J. Comp. Neurol. 501:231-242, 2007.

show abstract

ssDNA-dependent colocalization of adeno-associated virus Rep and herpes simplex virus ICP8 in nuclear replication domains

Cited by 26 publications

References 49 publications

hnRNP-U directly interacts with WT1 and modulates WT1 transcriptional activation

hnRNP-U directly interacts with WT1 and modulates WT1 transcriptional activation

Interactions Between AAV-2 and HSV-1: Implications for Hybrid Vector Design

Evidence for multiple signaling pathways in single squid olfactory receptor neurons

Contact Info

Product

Resources

About