Mapping Influenza-Induced Posttranslational Modifications on Histones from CD8+ T Cells
1
Department of Pediatrics, University of Tennessee Health Science Center, Memphis, TN 38163, USA
2
Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99354, USA
3
Children’s Foundation Research Institute, Memphis, TN 38105, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Italo Tempera
Viruses 2020, 12(12), 1409; https://doi.org/10.3390/v12121409
Received: 10 October 2020
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Revised: 23 November 2020
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Accepted: 2 December 2020
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Published: 8 December 2020
(This article belongs to the Special Issue Epigenetics and Viral Infection)
T cell function is determined by transcriptional networks that are regulated by epigenetic programming via posttranslational modifications (PTMs) to histone proteins and DNA. Bottom-up mass spectrometry (MS) can identify histone PTMs, whereas intact protein analysis by MS can detect species missed by bottom-up approaches. We used a novel approach of online two-dimensional liquid chromatography-tandem MS with high-resolution reversed-phase liquid chromatography (RPLC), alternating electron transfer dissociation (ETD) and collision-induced dissociation (CID) on precursor ions to maximize fragmentation of uniquely modified species. The first online RPLC separation sorted histone families, then RPLC or weak cation exchange hydrophilic interaction liquid chromatography (WCX-HILIC) separated species heavily clad in PTMs. Tentative identifications were assigned by matching proteoform masses to predicted theoretical masses that were verified with tandem MS. We used this innovative approach for histone-intact protein PTM mapping (HiPTMap) to identify and quantify proteoforms purified from CD8 T cells after in vivo influenza infection. Activation significantly altered PTMs following influenza infection, histone maps changed as T cells migrated to the site of infection, and T cells responding to secondary infections had significantly more transcription enhancing modifications. Thus, HiPTMap identified and quantified proteoforms and determined changes in CD8 T cell histone PTMs over the course of infection.
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Keywords:
epigenetic; histone; posttranslational modifications; T cells; influenza; top-down; mass spectrometry
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MDPI and ACS Style
Rezinciuc, S.; Tian, Z.; Wu, S.; Hengel, S.; Pasa-Tolic, L.; Smallwood, H.S. Mapping Influenza-Induced Posttranslational Modifications on Histones from CD8+ T Cells. Viruses 2020, 12, 1409. https://doi.org/10.3390/v12121409
AMA Style
Rezinciuc S, Tian Z, Wu S, Hengel S, Pasa-Tolic L, Smallwood HS. Mapping Influenza-Induced Posttranslational Modifications on Histones from CD8+ T Cells. Viruses. 2020; 12(12):1409. https://doi.org/10.3390/v12121409
Chicago/Turabian StyleRezinciuc, Svetlana, Zhixin Tian, Si Wu, Shawna Hengel, Ljiljana Pasa-Tolic, and Heather S. Smallwood. 2020. "Mapping Influenza-Induced Posttranslational Modifications on Histones from CD8+ T Cells" Viruses 12, no. 12: 1409. https://doi.org/10.3390/v12121409
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