We used pulse-labeling with the methionine analogue homopropargylglycine (HPG) to investigate spatiotemporal aspects of protein synthesis during herpes simplex virus (HSV) infection. revealed new insight into spatiotemporal aspects of protein localisation during contamination. A striking feature was the quick accumulation of newly synthesised proteins not only in a general nuclear pattern but additionally in newly forming sub-compartments represented by small discrete foci. These newly synthesised protein domains (NPDs) were similar in size and morphology to PML domains but were more numerous and whereas PML domains were progressively disrupted NPDs were progressively induced and persisted. Immediate-early proteins ICP4 and ICP0 were excluded from NPDs but using an ICP0 mutant defective in PML disruption we show a clear spatial relationship between NPDs and PML domains with NPDs frequently forming immediately adjacent and co-joining persisting PML domains. Further analysis of location of the chaperone Hsc70 exhibited that while NPDs created early in contamination without overt Hsc70 recruitment later in contamination Hsc70 showed pronounced recruitment frequently in a coat-like fashion around NPDs. Moreover while ICP4 and ICP0 were excluded from NPDs ICP22 showed selective recruitment. Our data show that NPDs symbolize early recruitment of host and viral de novo translated protein to unique structural entities which are precursors to the previously explained VICE domains involved in protein quality control in the nucleus and reveal new features from which we propose spatially linked platforms of newly synthesised protein processing after nuclear import. Author Summary All viruses reprogram infected cells for the synthesis modification and targeted localisation of virus-encoded and host proteins. Improvements in proteomics and mass spectrometry have provided broad insight into these processes but these methods have limited ability to investigate spatial aspects of infected Piragliatin cell protein synthesis and localisation. Here we provide the first statement using novel techniques in chemical biology including labeling newly synthesised proteins with chemically tagged amino acid precursors that enables subsequent biochemical analysis and spatial analysis by microscopy. Using these techniques we provide new insight into protein metabolism in herpes simplex virus infected cells which is not approachable by standard methods. We statement the formation of novel subnuclear domains termed NPDs (newly synthesised protein domains) with a Piragliatin spatial link to pre-existing nuclear PML domains and to previously explained domains involved in protein quality control. This work provides new insight into metabolic processes early after HSV contamination and demonstrates the considerable potential of these techniques to yield fundamental insight into virus contamination and virus-host interactions in any system. Introduction The manipulation of cellular metabolic processes during virus contamination promotes or tempers computer virus production and determines the outcome of infection not only at the cellular level but also e.g. acute versus long-term persistence latency reactivation and Piragliatin transmission [1]. With regard to infected cell protein metabolism as well as the regulated de novo synthesis of computer virus encoded proteins modulation of the host proteome is necessary for both contamination and host cell responses including modifications in protein turnover function and location [2]. Recent improvements in global proteomic methods and mass spectrometry methods have provided broad insight into the synthesis modification and degradation of viral and host proteins as contamination progresses [3-8]. These Piragliatin studies PDGFRA reveal alterations of cellular pathways including for example the remodeling of glycolytic and metabolic pathways [9] inflammatory and innate immune response factors [6 10 or nucleotide and RNA processing pathways [11]. However a complete understanding of infected cell protein metabolism requires a parallel approach to spatial aspects of global protein synthesis and transport dynamics and alterations in these processes during different stages of infection. Traditional analysis of proteins at steady-state using antibodies or fusion of genes to fluorescent.