TY - JOUR
T1 - Comprehensive Analyses of Intraviral Epstein-Barr Virus Protein-Protein Interactions Hint Central Role of BLRF2 in the Tegument Network
AU - Hara, Yuya
AU - Watanabe, Takahiro
AU - Yoshida, Masahiro
AU - Abdullah Al Masud, H. M.
AU - Kato, Hiromichi
AU - Kondo, Tomohiro
AU - Suzuki, Reiji
AU - Kurose, Shutaro
AU - Uddin, Md Kamal
AU - Arata, Masataka
AU - Miyagi, Shouhei
AU - Yanagi, Yusuke
AU - Sato, Yoshitaka
AU - Kimura, Hiroshi
AU - Murata, Takayuki
N1 - Funding Information:
We thank M. Miyata, S. Uchiayama, S. Kumagai, T. Kunogi, T. Kanda, H. Yoshiyama, Y. Narita, and T. Tsurumi for materials, technical assistance, and discussions. NanoBit screening, flow cytometry, and imaging analysis were performed at the Division for Medical Research Engineering, Nagoya University Graduate School of Medicine. This work was supported by grants-in-aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology (19K07580 to T.M., 21K15449 to T.W., 19K22560 and 20H03493 to H.K.), Japan Agency for Medical Research and Development (JP20wm0325012 to T.M.), the Takeda Science Foundation (to T.M.), and the Hori Sciences and Arts Foundation (to T.M., Y.S., and H.K.).
Publisher Copyright:
© 2022 American Society for Microbiology. All rights reserved.
PY - 2022/7
Y1 - 2022/7
N2 - Protein-protein interactions (PPIs) are crucial for various biological processes. Epstein-Barr virus (EBV) proteins typically form complexes, regulating the replication and persistence of the viral genome in human cells. However, the role of EBV protein complexes under physiological conditions remains unclear. In this study, we performed comprehensive analyses of EBV PPIs in living cells using the NanoBiT system. We identified 195 PPIs, many of which have not previously been reported. Computational analyses of these PPIs revealed that BLRF2, which is only found in gammaherpesviruses, is a central protein in the structural network of EBV tegument proteins. To characterize the role of BLRF2, we generated two BLRF2 knockout EBV clones using CRISPR/Cas9. BLRF2 knockout significantly decreased the production of infectious virus particles, which was partially restored by exogenous BLRF2 expression. In addition, self-association of BLRF2 protein was found, and mutation of the residues crucial for the self-association affected stability of the protein. Our data imply that BLRF2 is a tegument network hub that plays important roles in progeny virion maturation.
AB - Protein-protein interactions (PPIs) are crucial for various biological processes. Epstein-Barr virus (EBV) proteins typically form complexes, regulating the replication and persistence of the viral genome in human cells. However, the role of EBV protein complexes under physiological conditions remains unclear. In this study, we performed comprehensive analyses of EBV PPIs in living cells using the NanoBiT system. We identified 195 PPIs, many of which have not previously been reported. Computational analyses of these PPIs revealed that BLRF2, which is only found in gammaherpesviruses, is a central protein in the structural network of EBV tegument proteins. To characterize the role of BLRF2, we generated two BLRF2 knockout EBV clones using CRISPR/Cas9. BLRF2 knockout significantly decreased the production of infectious virus particles, which was partially restored by exogenous BLRF2 expression. In addition, self-association of BLRF2 protein was found, and mutation of the residues crucial for the self-association affected stability of the protein. Our data imply that BLRF2 is a tegument network hub that plays important roles in progeny virion maturation.
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U2 - 10.1128/jvi.00518-22
DO - 10.1128/jvi.00518-22
M3 - Article
C2 - 35862711
AN - SCOPUS:85135160416
SN - 0022-538X
VL - 96
JO - Journal of Virology
JF - Journal of Virology
IS - 14
ER -