Fluorescent TAP as a platform for virus-induced degradation of the antigenic peptide transporter

Magda Wąchalska , Małgorzata Graul , Patrique Praest , Rutger D. Luteijn , Aleksandra W. Babnis , Emmanuel J. H. J. Wiertz , Krystyna Bieńkowska-Szewczyk , Andrea D. Lipińska

Abstract

Transporter associated with antigen processing (TAP), a key player in the major histocompatibility complex class I-restricted antigen presentation, makes an attractive target for viruses that aim to escape the immune system. Mechanisms of TAP inhibition vary among virus species. Bovine herpesvirus 1 (BoHV-1) is unique in its ability to target TAP for proteasomal degradation following conformational arrest by the UL49.5 gene product. The exact mechanism of TAP removal still requires elucidation. For this purpose, a TAP-GFP (green fluorescent protein) fusion protein is instrumental, yet GFP-tagging may affect UL49.5-induced degradation. Therefore, we constructed a series of TAP-GFP variants using various linkers to obtain an optimal cellular fluorescent TAP platform. Mel JuSo (MJS) cells with CRISPR/Cas9 TAP1 or TAP2 knockouts were reconstituted with TAP-GFP constructs. Our results point towards a critical role of GFP localization on fluorescent properties of the fusion proteins and, in concert with the type of a linker, on the susceptibility to virally-induced inhibition and degradation. The fluorescent TAP platform was also used to re-evaluate TAP stability in the presence of other known viral TAP inhibitors, among which only UL49.5 was able to reduce TAP levels. Finally, we provide evidence that BoHV-1 UL49.5-induced TAP removal is p97-dependent, which indicates its degradation via endoplasmic reticulum-associated degradation (ERAD).
Author Magda Wąchalska (IFB)
Magda Wąchalska,,
- Intercollegiate Faculty of Biotechnology UG
, Małgorzata Graul (IFB / IB / LVMB)
Małgorzata Graul,,
- Laboratory of Virus Molecular Biology
, Patrique Praest
Patrique Praest,,
-
, Rutger D. Luteijn
Rutger D. Luteijn,,
-
, Aleksandra W. Babnis (IFB)
Aleksandra W. Babnis,,
- Intercollegiate Faculty of Biotechnology UG
, Emmanuel J. H. J. Wiertz
Emmanuel J. H. J. Wiertz,,
-
, Krystyna Bieńkowska-Szewczyk (IFB / IB / LVMB)
Krystyna Bieńkowska-Szewczyk,,
- Laboratory of Virus Molecular Biology
, Andrea D. Lipińska (IFB / IB / LVMB)
Andrea D. Lipińska,,
- Laboratory of Virus Molecular Biology
Journal seriesCells, ISSN 2073-4409, e-ISSN 2073-4409, (N/A 140 pkt)
Issue year2019
Vol8
No12
Pages1-20
Publication size in sheets0.95
Article number1590
Keywords in EnglishTAP-GFP, fluorescent TAP platform, antigen presentation, MHC I, immune evasion, BoHV-1 UL49.5
ASJC Classification1300 General Biochemistry, Genetics and Molecular Biology
DOIDOI:10.3390/cells8121590
URL https://www.mdpi.com/2073-4409/8/12/1590/pdf
Languageen angielski
LicenseJournal (articles only); published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)140
Score sourcejournalList
ScoreMinisterial score = 140.0, 05-03-2020, ArticleFromJournal
Publication indicators WoS Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2017 = 1.344; WoS Impact Factor: 2017 = 4.829 (2)
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