The KSHV portal protein ORF43 is essential for the production of infectious viral particles

Daniela (Dana) Dünn-Kittenplon; Inna Kalt; Jean Paul (Moshe) Lellouche; Ronit Sarid

doi:10.1016/j.virol.2019.01.028

The KSHV portal protein ORF43 is essential for the production of infectious viral particles

Daniela (Dana) Dünn-Kittenplon, Inna Kalt, Jean Paul (Moshe) Lellouche, Ronit Sarid

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

11 Scopus citations

Abstract

Herpesvirus capsid assembly involves cleavage and packaging of the viral genome. The Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 43 (orf43) encodes a putative portal protein. The portal complex functions as a gate through which DNA is packaged into the preformed procapsids, and is injected into the cell nucleus upon infection. The amino acid sequence of the portal proteins is conserved among herpesviruses. Here, we generated an antiserum to ORF43 and determined late expression kinetics of ORF43 along with its nuclear localization. We generated a recombinant KSHV mutant, which fails to express ORF43 (BAC16-ORF43-null). Assembled capsids were observed upon lytic induction of this virus; however, the released virions lacked viral DNA and thus could not establish infection. Ectopic expression of ORF43 rescued the ability to produce infectious particles. ORF43 antiserum and the recombinant ORF43-null virus can provide an experimental system for further studies of the portal functions and its interactions.

Original language	English
Pages (from-to)	205-215
Number of pages	11
Journal	Virology
Volume	529
DOIs	https://doi.org/10.1016/j.virol.2019.01.028
State	Published - Mar 2019

Bibliographical note

Publisher Copyright:
© 2019 Elsevier Inc.

Funding

We thank Dr. Julia Guez-Haddad for her valuable advice and help in bacterial expression and purification of ORF43 protein, Dr. Avi Jacobs and Dr. Irit Shoval for their assistance with confocal microscopy, Dr. Ayelet Atkins from the BIU TEM Unit, and Dr. Ilana Loinger for her advice on PCR settings. We also thank Anastasia Gelgor for her constructive advices and help. We gratefully acknowledge Prof. Jae Jung, Don Ganem, Shou Jiang Gao, Yan Yuan, Keiji Ueda and David Lukac for gifts of reagents. This work was supported by a grant from the Israel Science Foundation (Grant no. 1365/15 ).

Funders	Funder number
Israel Science Foundation	1365/15

Keywords

Kaposi's sarcoma-associated herpesvirus, KSHV
ORF43
Portal protein

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title = "The KSHV portal protein ORF43 is essential for the production of infectious viral particles",

abstract = "Herpesvirus capsid assembly involves cleavage and packaging of the viral genome. The Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 43 (orf43) encodes a putative portal protein. The portal complex functions as a gate through which DNA is packaged into the preformed procapsids, and is injected into the cell nucleus upon infection. The amino acid sequence of the portal proteins is conserved among herpesviruses. Here, we generated an antiserum to ORF43 and determined late expression kinetics of ORF43 along with its nuclear localization. We generated a recombinant KSHV mutant, which fails to express ORF43 (BAC16-ORF43-null). Assembled capsids were observed upon lytic induction of this virus; however, the released virions lacked viral DNA and thus could not establish infection. Ectopic expression of ORF43 rescued the ability to produce infectious particles. ORF43 antiserum and the recombinant ORF43-null virus can provide an experimental system for further studies of the portal functions and its interactions.",

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author = "D{\"u}nn-Kittenplon, {Daniela (Dana)} and Inna Kalt and Lellouche, {Jean Paul (Moshe)} and Ronit Sarid",

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doi = "10.1016/j.virol.2019.01.028",

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AU - Kalt, Inna

AU - Lellouche, Jean Paul (Moshe)

AU - Sarid, Ronit

PY - 2019/3

Y1 - 2019/3

N2 - Herpesvirus capsid assembly involves cleavage and packaging of the viral genome. The Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 43 (orf43) encodes a putative portal protein. The portal complex functions as a gate through which DNA is packaged into the preformed procapsids, and is injected into the cell nucleus upon infection. The amino acid sequence of the portal proteins is conserved among herpesviruses. Here, we generated an antiserum to ORF43 and determined late expression kinetics of ORF43 along with its nuclear localization. We generated a recombinant KSHV mutant, which fails to express ORF43 (BAC16-ORF43-null). Assembled capsids were observed upon lytic induction of this virus; however, the released virions lacked viral DNA and thus could not establish infection. Ectopic expression of ORF43 rescued the ability to produce infectious particles. ORF43 antiserum and the recombinant ORF43-null virus can provide an experimental system for further studies of the portal functions and its interactions.

AB - Herpesvirus capsid assembly involves cleavage and packaging of the viral genome. The Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 43 (orf43) encodes a putative portal protein. The portal complex functions as a gate through which DNA is packaged into the preformed procapsids, and is injected into the cell nucleus upon infection. The amino acid sequence of the portal proteins is conserved among herpesviruses. Here, we generated an antiserum to ORF43 and determined late expression kinetics of ORF43 along with its nuclear localization. We generated a recombinant KSHV mutant, which fails to express ORF43 (BAC16-ORF43-null). Assembled capsids were observed upon lytic induction of this virus; however, the released virions lacked viral DNA and thus could not establish infection. Ectopic expression of ORF43 rescued the ability to produce infectious particles. ORF43 antiserum and the recombinant ORF43-null virus can provide an experimental system for further studies of the portal functions and its interactions.

KW - Kaposi's sarcoma-associated herpesvirus, KSHV

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KW - Portal protein

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The KSHV portal protein ORF43 is essential for the production of infectious viral particles

Abstract

Bibliographical note

Funding

Keywords

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