Abstract
The organelles within a eukaryotic host are manipulated by viruses to support successful virus replication and spread of infection, yet the global impact of viral infection on host organelles is poorly understood. Integrating microscopy, subcellular fractionation, mass spectrometry, and functional analyses, we conducted a cell-wide study of organelles in primary fibroblasts throughout the time course of human cytomegalovirus (HCMV) infection. We used label-free and isobaric-labeling proteomics to characterize nearly 4,000 host and 100 viral proteins, then classified their specific subcellular locations over time using machine learning. We observed a global reorganization of proteins across the secretory pathway, plasma membrane, and mitochondria, including reorganization and processing of lysosomal proteins into distinct subpopulations and translocations of individual proteins between organelles at specific time points. We also demonstrate that MYO18A, an unconventional myosin that translocates from the plasma membrane to the viral assembly complex, is necessary for efficient HCMV replication. This study provides a comprehensive resource for understanding host and virus biology during HCMV pathogenesis.
🔬 Techniques
✨ Fluorophores
🧪 Sample Preparation
🔬 Cell Lines
🏭 Microscope Brands
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1
Cytoplasmic organelles undergo changes in shape, size, number, and organization following HCMV infection
A HCMV life cycle and the functional roles of organelles in viralnreplication. B Representative images of the structural reorganization of organelles in HCMVninfection (nu226517). Images acquired from...
Figure 2
Proteomic approach to define spatial-temporal changes in organelle proteins throughout HCMV infection
A Workflow to determine temporal changes in organelle proteins throughoutninfection. B Workflow to determine changes in the subcellular localization of proteinsnthroughout infection. C Data analysis t...
Figure 3
Temporal label-free proteomic analysis of organelles throughout HCMV infection
A Heatmap showing temporal changes of organelle proteins throughout infection.nOnly proteins with no missing values were displayed (2065 proteins).nHierarchical cluster determined five average cluster...
Figure 4
Protein subcellular localization assignment using organelle fractionation and alterations in the subcellular organization of lysosomal proteins
A 2-D representation of subcellular fractionation data using tSNE. The maps werenoverlaid with organelle markers (left) or assigned subcellular localizationsnusing machine learning (right). Sizes are ...
Figure 5
Temporal localizations and abundances of viral proteins throughout infection
A Heatmap of the subcellular localization of viral proteins and their relativenabundances throughout infection. Viral proteins were assigned localization tonperoxisomes (brown), mitochondria (red), ly...
Figure 6
Protein translocation upon HCMV infection
A Heatmap showing the number of translocation events at each time point ofninfection. Rows indicate location in uninfected samples, and columns indicatenlocalization at a specific time point of infect...
Figure 7
MYO18A translocates to the viral Assembly Complex and is important for viral production
A 2-D visualization for MYO18A gradient profile and its relation to othernorganelle-assigned proteins. B Gradient profile for MYO18A, MYO18A-associated host proteins (MYH9/10 andnGOLGB1), and markers ...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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