Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection
<p>Abstract</p> <p>A large quantitative study was carried out to compare the proteome of respiratory syncytial virus (RSV) infected versus uninfected cells in order to determine novel pathways regulated during viral infection. RSV infected and mock-infected HEp2 cells were lysed an...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2011-09-01
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| Series: | Virology Journal |
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| Online Access: | http://www.virologyj.com/content/8/1/442 |
| _version_ | 1828219450700070912 |
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| author | Altun Mikael Kramer Holger B Wright Cynthia Ternette Nicola Kessler Benedikt M |
| author_facet | Altun Mikael Kramer Holger B Wright Cynthia Ternette Nicola Kessler Benedikt M |
| author_sort | Altun Mikael |
| collection | DOAJ |
| description | <p>Abstract</p> <p>A large quantitative study was carried out to compare the proteome of respiratory syncytial virus (RSV) infected versus uninfected cells in order to determine novel pathways regulated during viral infection. RSV infected and mock-infected HEp2 cells were lysed and proteins separated by preparative isoelectric focussing using offgel fractionation. Following tryptic digestion, purified peptides were characterized using label-free quantitative expression profiling by nano-ultra performance liquid chromatography coupled to electrospray ionisation mass spectrometry with collision energy ramping for all-ion fragmentation (UPLC-MS<sup>E</sup>). A total of 1352 unique cellular proteins were identified and their abundance compared between infected and non-infected cells. Ingenuity pathway analysis revealed regulation of several central cellular metabolic and signalling pathways during infection. Selected proteins that were found regulated in RSV infected cells were screened by quantitative real-time PCR for their regulation on the transcriptional level. Synthesis of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) mRNAs were found to be highly induced upon RSV infection in a time dependent manner. Accordingly, IFIT3 protein levels accumulated during the time course of infection. In contrast, little variation was observed in XRN2 protein levels, but different forms were present in infected versus non-infected cells. This suggests a role of these proteins in viral infection, and analysis of their function will shed further light on mechanisms of RNA virus replication and the host cell defence machinery.</p> |
| first_indexed | 2024-04-12T16:15:50Z |
| format | Article |
| id | doaj.art-d92e63fd770f4f19abfb0dacbb64171a |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-04-12T16:15:50Z |
| publishDate | 2011-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-d92e63fd770f4f19abfb0dacbb64171a2022-12-22T03:25:43ZengBMCVirology Journal1743-422X2011-09-018144210.1186/1743-422X-8-442Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infectionAltun MikaelKramer Holger BWright CynthiaTernette NicolaKessler Benedikt M<p>Abstract</p> <p>A large quantitative study was carried out to compare the proteome of respiratory syncytial virus (RSV) infected versus uninfected cells in order to determine novel pathways regulated during viral infection. RSV infected and mock-infected HEp2 cells were lysed and proteins separated by preparative isoelectric focussing using offgel fractionation. Following tryptic digestion, purified peptides were characterized using label-free quantitative expression profiling by nano-ultra performance liquid chromatography coupled to electrospray ionisation mass spectrometry with collision energy ramping for all-ion fragmentation (UPLC-MS<sup>E</sup>). A total of 1352 unique cellular proteins were identified and their abundance compared between infected and non-infected cells. Ingenuity pathway analysis revealed regulation of several central cellular metabolic and signalling pathways during infection. Selected proteins that were found regulated in RSV infected cells were screened by quantitative real-time PCR for their regulation on the transcriptional level. Synthesis of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) mRNAs were found to be highly induced upon RSV infection in a time dependent manner. Accordingly, IFIT3 protein levels accumulated during the time course of infection. In contrast, little variation was observed in XRN2 protein levels, but different forms were present in infected versus non-infected cells. This suggests a role of these proteins in viral infection, and analysis of their function will shed further light on mechanisms of RNA virus replication and the host cell defence machinery.</p>http://www.virologyj.com/content/8/1/442Respiratory syncytial viruslabel-free quantitative proteomicsmass spectrometryIFIT3XRN2 |
| spellingShingle | Altun Mikael Kramer Holger B Wright Cynthia Ternette Nicola Kessler Benedikt M Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection Virology Journal Respiratory syncytial virus label-free quantitative proteomics mass spectrometry IFIT3 XRN2 |
| title | Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection |
| title_full | Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection |
| title_fullStr | Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection |
| title_full_unstemmed | Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection |
| title_short | Label-free quantitative proteomics reveals regulation of interferon-induced protein with tetratricopeptide repeats 3 (IFIT3) and 5'-3'-exoribonuclease 2 (XRN2) during respiratory syncytial virus infection |
| title_sort | label free quantitative proteomics reveals regulation of interferon induced protein with tetratricopeptide repeats 3 ifit3 and 5 3 exoribonuclease 2 xrn2 during respiratory syncytial virus infection |
| topic | Respiratory syncytial virus label-free quantitative proteomics mass spectrometry IFIT3 XRN2 |
| url | http://www.virologyj.com/content/8/1/442 |
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