Definition of herpes simplex virus helper functions for the replication of adeno-associated virus type 5

Stutika, Catrin; Hüser, Daniela; Weger, Stefan; Rutz, Natalja; Heßler, Melanie; Heilbronn, Regine

doi:10.1099/vir.0.000034

Cited by 9 publications

(6 citation statements)

References 47 publications

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“…Although most studies have focused on HSV-1 and AAV2 coinfections, it is likely that the same HSV-1 factors also support the replication of other AAV serotypes. A study by Stutika et al analyzed HSV-1 helper factors supporting AAV5 replication and concluded that the previously revealed helper functions for AAV2 also apply to AAV5 [ 150 ]. Since AAV5 is the most distantly related strain of AAV and was, in contrast to AAV1–4, directly isolated from human tissue, it is likely that the same HSV-1 factors, which support AAV2 and AAV5 replication, also support other AAV serotypes [ 151 , 152 ].…”

Section: Helper Viruses and Aavmentioning

confidence: 99%

The Interplay between Adeno-Associated Virus and Its Helper Viruses

Meier

Fraefel

Seyffert

2020

Viruses

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The adeno-associated virus (AAV) is a small, nonpathogenic parvovirus, which depends on helper factors to replicate. Those helper factors can be provided by coinfecting helper viruses such as adenoviruses, herpesviruses, or papillomaviruses. We review the basic biology of AAV and its most-studied helper viruses, adenovirus type 5 (AdV5) and herpes simplex virus type 1 (HSV-1). We further outline the direct and indirect interactions of AAV with those and additional helper viruses.

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Section: Helper Viruses and Aavmentioning

confidence: 99%

The Interplay between Adeno-Associated Virus and Its Helper Viruses

Meier

Fraefel

Seyffert

2020

Viruses

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“…HeLa cells seeded at a density of 30% were infected 20 h later with AAV2 wild-type or AAV-A1 mutant (multiplicity of infection [MOI] 250) and/or Ad2 (MOI 25). Transfection of HeLa cells with AAV plasmids was performed as described previously (21). Infection with Ad2 (MOI 25) was performed 16 h post- …”

Section: Methodsmentioning

confidence: 99%

A Comprehensive RNA Sequencing Analysis of the Adeno-Associated Virus (AAV) Type 2 Transcriptome Reveals Novel AAV Transcripts, Splice Variants, and Derived Proteins

Stutika

Gogol-Döring

Botschen

et al. 2016

J Virol

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Adeno-associated virus (AAV) is recognized for its bipartite life cycle with productive replication dependent on coinfection with adenovirus (Ad) and AAV latency being established in the absence of a helper virus. The shift from latent to Ad-dependent AAV replication is mostly regulated at the transcriptional level. The current AAV transcription map displays highly expressed transcripts as found upon coinfection with Ad. So far, AAV transcripts have only been characterized on the plus strand of the AAV single-stranded DNA genome. The AAV minus strand is assumed not to be transcribed. Here, we apply Illumina-based RNA sequencing (RNA-Seq) to characterize the entire AAV2 transcriptome in the absence or presence of Ad. We find known and identify novel AAV transcripts, including additional splice variants, the most abundant of which leads to expression of a novel 18-kDa Rep/VP fusion protein. Furthermore, we identify for the first time transcription on the AAV minus strand with clustered reads upstream of the p5 promoter, confirmed by 5' rapid amplification of cDNA ends and RNase protection assays. The p5 promoter displays considerable activity in both directions, a finding indicative of divergent transcription. Upon infection with AAV alone, low-level transcription of both AAV strands is detectable and is strongly stimulated upon coinfection with Ad. A deno-associated viruses (AAV) are helper-dependent members of the parvovirus group that require coinfection with an unrelated helper virus, particularly adenovirus (Ad) for productive replication (1). Despite of several identified AAV serotypes most research has been done with prototype AAV type 2. The AAV2 genome consists of a linear, single-stranded DNA of 4.7 kb. Both ends carry identical inverted terminal repeats (ITR) of 145 bp (2), which flank the two major open reading frames (ORF), called rep and cap. The rep ORF codes for four nonstructural proteins, Rep78 and a C-terminally spliced variant, Rep68. In addition, N-terminally truncated versions thereof are expressed, called Rep52 and Rep40, respectively. The Rep proteins are required as regulators for various steps of the AAV life cycle. AAV cap harbors the ORF for the capsid proteins (VP1 to -3) and a separate ORF for the assembly-activating protein (AAP) (3).The current knowledge of AAV transcription dates back to early research in the 1980s when the three AAV2 promoters, p5, p19, or p40 and major transcripts derived thereof were characterized (4, 5). All AAV transcripts identified since then map to only one DNA plus strand (6), which led to the assumption that the complementary AAV minus strand was not transcribed. Furthermore, spliced AAV transcripts were identified displaying a single, shared intron located in the center of the AAV2 genome (4, 7). Initially, a single splice donor site at AAV nucleotide 1906 and a single splice acceptor site located at nucleotide (nt) 2228 were described. Later, an alternative splice acceptor site at nt 2201 was detected (8), allowing the expression of VP1 protein ...

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“…Light green circles mark GFP-labeled structures by H8 tracer at both 36 hpi and 48 hpi; dark green circles mark GFP-labeled structures at 48 hpi can be directly used by AAV for its efficient replication and packaging. The required HSV genes for AAV production include UL5, UL8, UL9, UL29, UL30, UL42, and UL52, all of which are components of the HSV core replication system [35,36].…”

Section: Discussionmentioning

confidence: 99%

High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy

Ying

Han

et al. 2020

Mol Brain

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Neurotropic viral transsynaptic tracing is an increasingly powerful technique for dissecting the structure and function of neural circuits. Herpes simplex virus type 1 strain H129 has been widely used as an anterograde tracer. However, HSV tracers still have several shortcomings, including high toxicity, low sensitivity and non-specific retrograde labeling. Here, we aimed to construct high-brightness HSV anterograde tracers by increasing the expression of exogenous genes carried by H129 viruses. Using a Trojan horse-like strategy, a HSV/AAV (adenoassociated virus) chimaera termed H8 was generated to enhance the expression of a fluorescent marker. In vitro and in vivo assays showed that the exogenous gene was efficiently replicated and amplified by the synergism of the HSV vector and introduced AAV replication system. H8 reporting fluorescence was brighter than that of currently available H129 tracers, and H8 could be used for fast and effective anterograde tracing without additional immunostaining. These results indicated that foreign gene expression in HSV tracers could be enhanced by integrating HSV with AAV replication system. This approach may be useful as a general enhanced expression strategy for HSV-based tracing tools or gene delivery vectors.

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Definition of herpes simplex virus helper functions for the replication of adeno-associated virus type 5

Cited by 9 publications

References 47 publications

The Interplay between Adeno-Associated Virus and Its Helper Viruses

The Interplay between Adeno-Associated Virus and Its Helper Viruses

A Comprehensive RNA Sequencing Analysis of the Adeno-Associated Virus (AAV) Type 2 Transcriptome Reveals Novel AAV Transcripts, Splice Variants, and Derived Proteins

High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy

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