1 Cellular protein TTC4 and its cofactor HSP90 are pro-viral for bovine herpesvirus 1

Beth H  Thompson; Colin Sharp; Inga Dry; Robert Dalziel; Eleanor Gaunt

doi:10.1016/j.virusres.2022.198927

1 Cellular protein TTC4 and its cofactor HSP90 are pro-viral for bovine herpesvirus 1

Beth H Thompson, Colin Sharp, Inga Dry, Robert Dalziel, Eleanor Gaunt^*

^*Corresponding author for this work

Royal (Dick) School of Veterinary Studies

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

Abstract / Description of output

Bovine Herpesvirus Type 1 (BoHV-1) infection causes infectious bovine rhinotracheitis and genital disease in cattle, with significant economic and welfare impacts. However, the role of cellular host factors during viral replication remains poorly characterised. A previously performed genome-wide CRISPR knockout screen identified pro- and antiviral host factors acting during BoHV-1 replication. Herein we validate a pro-viral role for a candidate from this screen: the cellular protein tetracopeptide repeat protein 4 (TTC4). We show that TTC4 transcript production is upregulated during BoHV-1 infection. Depletion of TTC4 protein impairs BoHV-1 protein production but does not reduce production of infectious virions, whereas overexpression of exogenous TTC4 results in a significant increase in production of infectious BoHV-1 virions. TTC4 itself is poorly characterized (especially in the context of virus infection), but is a known co-chaperone of heat shock protein 90 (HSP90). HSP90 has a well-characterized pro-viral role during the replication of diverse herpesviruses, and we therefore hypothesized that HSP90 is also pro-viral for BoHV-1. Drug-mediated inhibition of HSP90 using geldanamycin at sub-cytotoxic concentrations inhibited both BoHV-1 protein production and viral genome replication, indicating a pro-viral role for HSP90 during BoHV-1 infection. Our data demonstrates pro-viral roles for both TTC4 and HSP90 during BoHV-1 replication; possibly, interactions between these two proteins are required for optimal BoHV-1 replication, or the two proteins may have independent pro-viral roles.

Original language	English
Article number	198927
Journal	Virus Research
Volume	321
Early online date	10 Sept 2022
DOIs	https://doi.org/10.1016/j.virusres.2022.198927
Publication status	Published - Nov 2022

Keywords / Materials (for Non-textual outputs)

Bovine herpesvirus 1
tetracopeptide repeat protein 4 (TTC4)
heat shock protein 90 (HSP90)
geldanamycin

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10.1016/j.virusres.2022.198927

TTC4_Manuscript_changes_accepted_5Sep22Accepted author manuscript, 0.98 MBLicence: Creative Commons: Attribution (CC-BY)

Cite this

@article{d512c6ba7e3c4553905a24296a60e36a,

title = "1 Cellular protein TTC4 and its cofactor HSP90 are pro-viral for bovine herpesvirus 1",

abstract = "Bovine Herpesvirus Type 1 (BoHV-1) infection causes infectious bovine rhinotracheitis and genital disease in cattle, with significant economic and welfare impacts. However, the role of cellular host factors during viral replication remains poorly characterised. A previously performed genome-wide CRISPR knockout screen identified pro- and antiviral host factors acting during BoHV-1 replication. Herein we validate a pro-viral role for a candidate from this screen: the cellular protein tetracopeptide repeat protein 4 (TTC4). We show that TTC4 transcript production is upregulated during BoHV-1 infection. Depletion of TTC4 protein impairs BoHV-1 protein production but does not reduce production of infectious virions, whereas overexpression of exogenous TTC4 results in a significant increase in production of infectious BoHV-1 virions. TTC4 itself is poorly characterized (especially in the context of virus infection), but is a known co-chaperone of heat shock protein 90 (HSP90). HSP90 has a well-characterized pro-viral role during the replication of diverse herpesviruses, and we therefore hypothesized that HSP90 is also pro-viral for BoHV-1. Drug-mediated inhibition of HSP90 using geldanamycin at sub-cytotoxic concentrations inhibited both BoHV-1 protein production and viral genome replication, indicating a pro-viral role for HSP90 during BoHV-1 infection. Our data demonstrates pro-viral roles for both TTC4 and HSP90 during BoHV-1 replication; possibly, interactions between these two proteins are required for optimal BoHV-1 replication, or the two proteins may have independent pro-viral roles. ",

keywords = "Bovine herpesvirus 1, tetracopeptide repeat protein 4 (TTC4), heat shock protein 90 (HSP90), geldanamycin",

author = "Thompson, {Beth H} and Colin Sharp and Inga Dry and Robert Dalziel and Eleanor Gaunt",

note = "Funding Information: We are indebted to the staff in the Central Services Unit and Bioimaging facilities in the Roslin Institute, University of Edinburgh. ERG, RGD, CPS and IRD are supported by a BBSRC Institute Strategic Programme grant (BB/P013740/1). ERG is also supported by a Wellcome Trust/ Royal Society Sir Henry Dale Fellowship (211222_Z_18_Z). RGD was supported by BBSRC grant BB/P013740/1. BT was supported by a MSD animal health research bursary, a Microbiology Society Harry Smith Vacation studentship (GA002550) and Wellcome Trust ISSF3 funding. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. For the purpose of open access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. We are grateful to Dr Wenfang S Tan (The Roslin Institute) for provision of reagents. Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = nov,

doi = "10.1016/j.virusres.2022.198927",

language = "English",

volume = "321",

journal = "Virus Research",

issn = "0168-1702",

publisher = "Elsevier",

}

TY - JOUR

T1 - 1 Cellular protein TTC4 and its cofactor HSP90 are pro-viral for bovine herpesvirus 1

AU - Thompson, Beth H

AU - Sharp, Colin

AU - Dry, Inga

AU - Dalziel, Robert

AU - Gaunt, Eleanor

N1 - Funding Information: We are indebted to the staff in the Central Services Unit and Bioimaging facilities in the Roslin Institute, University of Edinburgh. ERG, RGD, CPS and IRD are supported by a BBSRC Institute Strategic Programme grant (BB/P013740/1). ERG is also supported by a Wellcome Trust/ Royal Society Sir Henry Dale Fellowship (211222_Z_18_Z). RGD was supported by BBSRC grant BB/P013740/1. BT was supported by a MSD animal health research bursary, a Microbiology Society Harry Smith Vacation studentship (GA002550) and Wellcome Trust ISSF3 funding. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. For the purpose of open access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. We are grateful to Dr Wenfang S Tan (The Roslin Institute) for provision of reagents. Publisher Copyright: © 2022 The Author(s)

PY - 2022/11

Y1 - 2022/11

N2 - Bovine Herpesvirus Type 1 (BoHV-1) infection causes infectious bovine rhinotracheitis and genital disease in cattle, with significant economic and welfare impacts. However, the role of cellular host factors during viral replication remains poorly characterised. A previously performed genome-wide CRISPR knockout screen identified pro- and antiviral host factors acting during BoHV-1 replication. Herein we validate a pro-viral role for a candidate from this screen: the cellular protein tetracopeptide repeat protein 4 (TTC4). We show that TTC4 transcript production is upregulated during BoHV-1 infection. Depletion of TTC4 protein impairs BoHV-1 protein production but does not reduce production of infectious virions, whereas overexpression of exogenous TTC4 results in a significant increase in production of infectious BoHV-1 virions. TTC4 itself is poorly characterized (especially in the context of virus infection), but is a known co-chaperone of heat shock protein 90 (HSP90). HSP90 has a well-characterized pro-viral role during the replication of diverse herpesviruses, and we therefore hypothesized that HSP90 is also pro-viral for BoHV-1. Drug-mediated inhibition of HSP90 using geldanamycin at sub-cytotoxic concentrations inhibited both BoHV-1 protein production and viral genome replication, indicating a pro-viral role for HSP90 during BoHV-1 infection. Our data demonstrates pro-viral roles for both TTC4 and HSP90 during BoHV-1 replication; possibly, interactions between these two proteins are required for optimal BoHV-1 replication, or the two proteins may have independent pro-viral roles.

AB - Bovine Herpesvirus Type 1 (BoHV-1) infection causes infectious bovine rhinotracheitis and genital disease in cattle, with significant economic and welfare impacts. However, the role of cellular host factors during viral replication remains poorly characterised. A previously performed genome-wide CRISPR knockout screen identified pro- and antiviral host factors acting during BoHV-1 replication. Herein we validate a pro-viral role for a candidate from this screen: the cellular protein tetracopeptide repeat protein 4 (TTC4). We show that TTC4 transcript production is upregulated during BoHV-1 infection. Depletion of TTC4 protein impairs BoHV-1 protein production but does not reduce production of infectious virions, whereas overexpression of exogenous TTC4 results in a significant increase in production of infectious BoHV-1 virions. TTC4 itself is poorly characterized (especially in the context of virus infection), but is a known co-chaperone of heat shock protein 90 (HSP90). HSP90 has a well-characterized pro-viral role during the replication of diverse herpesviruses, and we therefore hypothesized that HSP90 is also pro-viral for BoHV-1. Drug-mediated inhibition of HSP90 using geldanamycin at sub-cytotoxic concentrations inhibited both BoHV-1 protein production and viral genome replication, indicating a pro-viral role for HSP90 during BoHV-1 infection. Our data demonstrates pro-viral roles for both TTC4 and HSP90 during BoHV-1 replication; possibly, interactions between these two proteins are required for optimal BoHV-1 replication, or the two proteins may have independent pro-viral roles.

KW - Bovine herpesvirus 1

KW - tetracopeptide repeat protein 4 (TTC4)

KW - heat shock protein 90 (HSP90)

KW - geldanamycin

U2 - 10.1016/j.virusres.2022.198927

DO - 10.1016/j.virusres.2022.198927

M3 - Article

SN - 0168-1702

VL - 321

JO - Virus Research

JF - Virus Research

M1 - 198927

ER -

1 Cellular protein TTC4 and its cofactor HSP90 are pro-viral for bovine herpesvirus 1

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