A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion

Mamadou Amadou Diallo; Sébastien Pirotte; Yunlong Hu; Léa Morvan; Krzysztof Rakus; Nicolás M. Suárez; Lee Potsang; Hisao Saneyoshi; Yan Xu; Andrew J. Davison; Peter Tompa; Joel L. Sussman; Alain Vanderplasschen

doi:10.1093/nar/gkac761

A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion

Mamadou Amadou Diallo, Sébastien Pirotte, Yunlong Hu, Léa Morvan, Krzysztof Rakus, Nicolás M. Suárez, Lee Potsang, Hisao Saneyoshi, Yan Xu, Andrew J. Davison, Peter Tompa, Joel L. Sussman, Alain Vanderplasschen^*

^*Corresponding author for this work

Department of Chemical and Structural Biology

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

Zalpha (Zα) domains bind to left-handed Z-DNA and Z-RNA. The Zα domain protein family includes cellular (ADAR1, ZBP1 and PKZ) and viral (vaccinia virus E3 and cyprinid herpesvirus 3 (CyHV-3) ORF112) proteins. We studied CyHV-3 ORF112, which contains an intrinsically disordered region and a Zα domain. Genome editing of CyHV-3 indicated that the expression of only the Zα domain of ORF112 was sufficient for normal viral replication in cell culture and virulence in carp. In contrast, its deletion was lethal for the virus. These observations revealed the potential of the CyHV-3 model as a unique platform to compare the exchangeability of Zα domains expressed alone in living cells. Attempts to rescue the ORF112 deletion by a broad spectrum of cellular, viral, and artificial Zα domains showed that only those expressing Z-binding activity, the capacity to induce liquid-liquid phase separation (LLPS), and A-to-Z conversion, could rescue viral replication. For the first time, this study reports the ability of some Zα domains to induce LLPS and supports the biological relevance of dsRNA A-to-Z conversion mediated by Zα domains. This study expands the functional diversity of Zα domains and stimulates new hypotheses concerning the mechanisms of action of proteins containing Zα domains.

Original language	English
Pages (from-to)	806-830
Number of pages	25
Journal	Nucleic Acids Research
Volume	51
Issue number	2
Early online date	22 Sept 2022
DOIs	https://doi.org/10.1093/nar/gkac761
Publication status	Published - 25 Jan 2023

Bibliographical note

S.P. and L.M. are research fellows of the FNRS. Y.H. is a research fellow of the Chinese Scholarship Council. The authors are grateful to the GIGA-Cell imaging platform of the University of Liège for its technical support. A.V. would like to express his gratitude to Isabelle Vierendeels for being such great source of inspiration for this project. This manuscript is dedicated to the memory of Dr Alekos Athanasiadis (Instituto Gulbenkian de Ciência, Portugal) who loved studying the biological functions of Zα domains.
Funding:University of Liège [ARC15/19-12 to A.V.]; Fonds National Belge de la Recherche Scientifique (FNRS) [CDR J.0094.15 and PDR T.0241.19 to A.V.]; Welbio [WELBIO-CR-2022 A-14 12 to A.V.]; Walloon Region [ERANets NucNanoFish 2010052 to A.V.]; Medical Research Council [MC_UU_12014/3 to A.J.D.]; Vrije Universiteit Brussel [Spearhead grant SRP51 to P.T.]; European Commission [EC H2020-WIDESPREAD-2020-5 Twinning grant PhasAge, 952334, and EC H2020-MSCA-RISE Action grant IDPfun 778247 to P.T.]; Japan Society for the promotion of science Kakenhi [21H02081 to Y.X.]. Funding for open access charge: University of Liège (Belgium).
Publisher Copyright:
© 2023 The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

All Science Journal Classification (ASJC) codes

Genetics

Access to Document

10.1093/nar/gkac761Licence: CC BY

Cite this

Diallo, M. A., Pirotte, S., Hu, Y., Morvan, L., Rakus, K., Suárez, N. M., Potsang, L., Saneyoshi, H., Xu, Y., Davison, A. J., Tompa, P., Sussman, J. L., & Vanderplasschen, A. (2023). A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion. Nucleic Acids Research, 51(2), 806-830. https://doi.org/10.1093/nar/gkac761

@article{c9499e9e68ab41c5a14f208572d6168d,

title = "A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion",

abstract = "Zalpha (Zα) domains bind to left-handed Z-DNA and Z-RNA. The Zα domain protein family includes cellular (ADAR1, ZBP1 and PKZ) and viral (vaccinia virus E3 and cyprinid herpesvirus 3 (CyHV-3) ORF112) proteins. We studied CyHV-3 ORF112, which contains an intrinsically disordered region and a Zα domain. Genome editing of CyHV-3 indicated that the expression of only the Zα domain of ORF112 was sufficient for normal viral replication in cell culture and virulence in carp. In contrast, its deletion was lethal for the virus. These observations revealed the potential of the CyHV-3 model as a unique platform to compare the exchangeability of Zα domains expressed alone in living cells. Attempts to rescue the ORF112 deletion by a broad spectrum of cellular, viral, and artificial Zα domains showed that only those expressing Z-binding activity, the capacity to induce liquid-liquid phase separation (LLPS), and A-to-Z conversion, could rescue viral replication. For the first time, this study reports the ability of some Zα domains to induce LLPS and supports the biological relevance of dsRNA A-to-Z conversion mediated by Zα domains. This study expands the functional diversity of Zα domains and stimulates new hypotheses concerning the mechanisms of action of proteins containing Zα domains.",

author = "Diallo, {Mamadou Amadou} and S{\'e}bastien Pirotte and Yunlong Hu and L{\'e}a Morvan and Krzysztof Rakus and Su{\'a}rez, {Nicol{\'a}s M.} and Lee Potsang and Hisao Saneyoshi and Yan Xu and Davison, {Andrew J.} and Peter Tompa and Sussman, {Joel L.} and Alain Vanderplasschen",

note = "S.P. and L.M. are research fellows of the FNRS. Y.H. is a research fellow of the Chinese Scholarship Council. The authors are grateful to the GIGA-Cell imaging platform of the University of Li{\`e}ge for its technical support. A.V. would like to express his gratitude to Isabelle Vierendeels for being such great source of inspiration for this project. This manuscript is dedicated to the memory of Dr Alekos Athanasiadis (Instituto Gulbenkian de Ci{\^e}ncia, Portugal) who loved studying the biological functions of Zα domains. Funding:University of Li{\`e}ge [ARC15/19-12 to A.V.]; Fonds National Belge de la Recherche Scientifique (FNRS) [CDR J.0094.15 and PDR T.0241.19 to A.V.]; Welbio [WELBIO-CR-2022 A-14 12 to A.V.]; Walloon Region [ERANets NucNanoFish 2010052 to A.V.]; Medical Research Council [MC_UU_12014/3 to A.J.D.]; Vrije Universiteit Brussel [Spearhead grant SRP51 to P.T.]; European Commission [EC H2020-WIDESPREAD-2020-5 Twinning grant PhasAge, 952334, and EC H2020-MSCA-RISE Action grant IDPfun 778247 to P.T.]; Japan Society for the promotion of science Kakenhi [21H02081 to Y.X.]. Funding for open access charge: University of Li{\`e}ge (Belgium). Publisher Copyright: {\textcopyright} 2023 The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.",

year = "2023",

month = jan,

day = "25",

doi = "10.1093/nar/gkac761",

language = "English",

volume = "51",

pages = "806--830",

journal = "Nucleic Acids Research",

issn = "0305-1048",

publisher = "Oxford University Press",

number = "2",

}

Diallo, MA, Pirotte, S, Hu, Y, Morvan, L, Rakus, K, Suárez, NM, Potsang, L, Saneyoshi, H, Xu, Y, Davison, AJ, Tompa, P, Sussman, JL & Vanderplasschen, A 2023, 'A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion', Nucleic Acids Research, vol. 51, no. 2, pp. 806-830. https://doi.org/10.1093/nar/gkac761

TY - JOUR

T1 - A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion

AU - Diallo, Mamadou Amadou

AU - Pirotte, Sébastien

AU - Hu, Yunlong

AU - Morvan, Léa

AU - Rakus, Krzysztof

AU - Suárez, Nicolás M.

AU - Potsang, Lee

AU - Saneyoshi, Hisao

AU - Xu, Yan

AU - Davison, Andrew J.

AU - Tompa, Peter

AU - Sussman, Joel L.

AU - Vanderplasschen, Alain

N1 - S.P. and L.M. are research fellows of the FNRS. Y.H. is a research fellow of the Chinese Scholarship Council. The authors are grateful to the GIGA-Cell imaging platform of the University of Liège for its technical support. A.V. would like to express his gratitude to Isabelle Vierendeels for being such great source of inspiration for this project. This manuscript is dedicated to the memory of Dr Alekos Athanasiadis (Instituto Gulbenkian de Ciência, Portugal) who loved studying the biological functions of Zα domains. Funding:University of Liège [ARC15/19-12 to A.V.]; Fonds National Belge de la Recherche Scientifique (FNRS) [CDR J.0094.15 and PDR T.0241.19 to A.V.]; Welbio [WELBIO-CR-2022 A-14 12 to A.V.]; Walloon Region [ERANets NucNanoFish 2010052 to A.V.]; Medical Research Council [MC_UU_12014/3 to A.J.D.]; Vrije Universiteit Brussel [Spearhead grant SRP51 to P.T.]; European Commission [EC H2020-WIDESPREAD-2020-5 Twinning grant PhasAge, 952334, and EC H2020-MSCA-RISE Action grant IDPfun 778247 to P.T.]; Japan Society for the promotion of science Kakenhi [21H02081 to Y.X.]. Funding for open access charge: University of Liège (Belgium). Publisher Copyright: © 2023 The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2023/1/25

Y1 - 2023/1/25

N2 - Zalpha (Zα) domains bind to left-handed Z-DNA and Z-RNA. The Zα domain protein family includes cellular (ADAR1, ZBP1 and PKZ) and viral (vaccinia virus E3 and cyprinid herpesvirus 3 (CyHV-3) ORF112) proteins. We studied CyHV-3 ORF112, which contains an intrinsically disordered region and a Zα domain. Genome editing of CyHV-3 indicated that the expression of only the Zα domain of ORF112 was sufficient for normal viral replication in cell culture and virulence in carp. In contrast, its deletion was lethal for the virus. These observations revealed the potential of the CyHV-3 model as a unique platform to compare the exchangeability of Zα domains expressed alone in living cells. Attempts to rescue the ORF112 deletion by a broad spectrum of cellular, viral, and artificial Zα domains showed that only those expressing Z-binding activity, the capacity to induce liquid-liquid phase separation (LLPS), and A-to-Z conversion, could rescue viral replication. For the first time, this study reports the ability of some Zα domains to induce LLPS and supports the biological relevance of dsRNA A-to-Z conversion mediated by Zα domains. This study expands the functional diversity of Zα domains and stimulates new hypotheses concerning the mechanisms of action of proteins containing Zα domains.

AB - Zalpha (Zα) domains bind to left-handed Z-DNA and Z-RNA. The Zα domain protein family includes cellular (ADAR1, ZBP1 and PKZ) and viral (vaccinia virus E3 and cyprinid herpesvirus 3 (CyHV-3) ORF112) proteins. We studied CyHV-3 ORF112, which contains an intrinsically disordered region and a Zα domain. Genome editing of CyHV-3 indicated that the expression of only the Zα domain of ORF112 was sufficient for normal viral replication in cell culture and virulence in carp. In contrast, its deletion was lethal for the virus. These observations revealed the potential of the CyHV-3 model as a unique platform to compare the exchangeability of Zα domains expressed alone in living cells. Attempts to rescue the ORF112 deletion by a broad spectrum of cellular, viral, and artificial Zα domains showed that only those expressing Z-binding activity, the capacity to induce liquid-liquid phase separation (LLPS), and A-to-Z conversion, could rescue viral replication. For the first time, this study reports the ability of some Zα domains to induce LLPS and supports the biological relevance of dsRNA A-to-Z conversion mediated by Zα domains. This study expands the functional diversity of Zα domains and stimulates new hypotheses concerning the mechanisms of action of proteins containing Zα domains.

UR - http://www.scopus.com/inward/record.url?scp=85149005781&partnerID=8YFLogxK

U2 - 10.1093/nar/gkac761

DO - 10.1093/nar/gkac761

M3 - Article

C2 - 36130731

AN - SCOPUS:85149005781

SN - 0305-1048

VL - 51

SP - 806

EP - 830

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 2

ER -

A fish herpesvirus highlights functional diversities among Zα domains related to phase separation induction and A-to-Z conversion

Abstract

Bibliographical note

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this