{"id":7185,"date":"2016-01-25T07:00:44","date_gmt":"2016-01-25T12:00:44","guid":{"rendered":"http:\/\/admin.acceleratingscience.com\/?p=7185"},"modified":"2016-04-29T16:53:11","modified_gmt":"2016-04-29T16:53:11","slug":"quantitative-temporal-viromics-investigates-hcmv","status":"publish","type":"post","link":"https:\/\/www.thermofisher.com\/blog\/proteomics\/quantitative-temporal-viromics-investigates-hcmv\/","title":{"rendered":"Quantitative Temporal Viromics investigates HCMV"},"content":{"rendered":"<p><span><img loading=\"lazy\" decoding=\"async\" src=\"http:\/\/admin.acceleratingscience.com\/proteomics\/wp-content\/uploads\/sites\/2\/2016\/01\/shutterstock_2082239531.jpg\" style=\"float: left;margin: 10px\" alt=\"Cytomegalovirus. Image: 306\/Shutterstock.com\" width=\"330\" height=\"164\" \/>Human cytomegalovirus (HCMV), like other herpes viruses, can remain latent for years without causing symptoms. If an infected individual experiences a decrease in immune function, symptoms can occur, and the disease can then be passed to others. <\/span><\/p>\n<p>In light of studying HCMV virus-host interaction, Weekes et al. pioneered a new method, quantitative temporal viromics (QTV), to look for key proteins involved in host defense.<sup>1<\/sup>&nbsp;The researchers used a combination of plasma membrane profiling with a study of whole-cell lysates (WCL). They quantified proteins from up to 10 samples using isobaric chemical tandem mass tags (TMT).<\/p>\n<p>Weekes et al. based the experimental design on their previously published method for isolating highly purified PM proteins for proteomic analysis.<sup>2<\/sup> By infecting primary human fetal foreskin fibroblasts (HFFFs) with HCMV strain, the team looked at changes in the expression of &nbsp;proteins from the plasma membrane across different time points.<\/p>\n<p><span>They used an <\/span><a href=\"http:\/\/www.thermoscientific.com\/en\/product\/orbitrap-elite-hybrid-ion-trap-orbitrap-mass-spectrometer.html\"><span>Orbitrap Elite mass spectrometer<\/span><\/a><span>&nbsp;(Thermo Scientific) for this analysis. From this, the researchers quantified 927 plasma membrane proteins and found 56% of proteins changed more than 2-fold and 33% more than 3-fold by hour 72 of infection. They also observed expected changes in 21 of 22 cellular proteins and detected 6 of 6 previously reported HCMV proteins at the plasma membrane. They determined that 5 proteins are virion envelope glycoproteins expressed late in infection. <\/span><\/p>\n<p><span class=\"thread\">The researchers also performed a temporal analysis of WCLs of HCMV infected HF<\/span>FFs. Looking at protein expression during infection combined with the total abundance of a given protein in the plasma membrane, the team chose an <a href=\"http:\/\/www.thermoscientific.com\/en\/product\/orbitrap-fusion-tribrid-mass-spectrometer.html\">Orbitrap Fusion Tribrid mass spectrometer<\/a>&nbsp;(Thermo Scientific). This further confirmed changes in 31 of 35 previously reported proteins.<\/p>\n<p><span>Taking a more in-depth look at QTV, this time utilizing 10-plex TMT and seven time points, the team quantified 1,184 plasma proteins and 7,491 cellular proteins.&nbsp;<\/span>In addition to finding a high degree of reproducibility among biological replicates,&nbsp;the team used a high-throughput Western blot analysis of cells infected with Toledo strain of HCMV to further confirm their results. They saw a strong association between protein changes in both experiments (p &lt; 0.0001).<\/p>\n<p>In conclusion, QTV helped identify temporal profiles of &nbsp;more than 80% of HCMV canonical genes and 14 non-canonical HCMV open reading frames. The researchers consider QTV a powerful method to study viral infection.<\/p>\n<p>&nbsp;<\/p>\n<p><strong>References<\/strong><\/p>\n<p><span>1. Weekes, M.P., et al. (2014) &ldquo;<\/span><span><a href=\"http:\/\/www.cell.com\/cell\/abstract\/S0092-8674(14)00594-7\">Quantitative temporal viromics: An approach to investigate host-pathogen interaction<\/a>,<\/span><span>&rdquo; Cell, 157 (pp. 1460&ndash;1472).<\/span><\/p>\n<p><span><span>2. Weekes, M.P., et al. &nbsp;(2012) &ldquo;<\/span><span><a href=\"http:\/\/www.ncbi.nlm.nih.gov\/pubmed\/22292497\">Proteomic plasma membrane profiling reveals an essential role for gp96 in the cell surface expression of LDLR family members, including the LDL receptor and LRP6<\/a>,<\/span><span>&rdquo; Journal of Proteome Research, 11 (pp. 1475&ndash;1484).<\/span><\/span><\/p>\n","protected":false},"excerpt":{"rendered":"<p>Human cytomegalovirus (HCMV), like other herpes viruses, can remain latent for years without causing symptoms. If an infected individual experiences a decrease in immune function, symptoms can occur, and the disease can then be passed to others. In light of studying HCMV virus-host interaction, Weekes et al. pioneered a new method, quantitative temporal viromics (QTV),<\/p>\n","protected":false},"author":13,"featured_media":7318,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"_acf_changed":false,"_monsterinsights_skip_tracking":false,"_genesis_hide_title":false,"_genesis_hide_breadcrumbs":false,"_genesis_hide_singular_image":false,"_genesis_hide_footer_widgets":false,"_genesis_custom_body_class":"","_genesis_custom_post_class":"","_genesis_layout":"","_jetpack_newsletter_access":"","_jetpack_dont_email_post_to_subs":false,"_jetpack_newsletter_tier_id":0,"_jetpack_memberships_contains_paywalled_content":false,"_jetpack_memberships_contains_paid_content":false,"footnotes":""},"categories":[17],"tags":[808],"division":[],"class_list":{"0":"post-7185","1":"post","2":"type-post","3":"status-publish","4":"format-standard","5":"has-post-thumbnail","7":"category-virology","8":"tag-hcmv","9":"entry"},"_selected_authors":"","_selected_reviewers":"","acf":[],"yoast_head":"<!-- This site is optimized 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