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Human cytomegalovirus UL24 and UL43 products participate in SAMHD1 subcellular localization

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Abstract

This report has analyzed the potential role of Human Cytomegalovirus (HCMV) UL24 and UL43 products in modulating the subcellular location of a host restriction factor, SAMHD1, in cells of human fibroblast origin. Recent studies have reported that the regulation of SAMHD1 is mediated by the HCMV UL97 product inside the nucleus, and by the CDK pathway when it is located in the cytoplasm of the infected cells but the viral gene products that may involve in cytosolic relocalization remain unknown yet. In the present report, we demonstrate that the HCMV UL24 product interacts with the SAMHD1 protein during infection based on mass spectrometry (MS) data and immunoprecipitation assay. The expression or depletion of the viral UL24 gene product did not affect the subcellular localization of SAMHD1 but when it coexpressed with the viral UL43 gene product, another member of the HCMV US22 family, induced the SAMHD1 cytosolic relocalization. Interestingly, the double deletion of viral UL24 and UL43 gene products impaired the cytosolic translocation and the SAMHD1 was accumulated in the nucleus of the infected cells, especially at the late stage post-infection. Our results provide evidence that the viral UL24 and UL43 gene products play a role in the SAMHD1 subcellular localization during HCMV infection.

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Abbreviations

HCMV AD169:

Human cytomegalovirus strain AD169

AGS:

Aicardi-Goutie'res syndrome

ATCC:

American Type Culture Collection

BAC:

Bacterial Artificial Chromosome

HCMV dd24:

Human cytomegalovirus with deleted UL24 gene

HCMV dd24–dd43:

Human cytomegalovirus with double deleted UL24 and UL43 genes

CD81:

Cluster of Differentiation 81

CDK1:

Cyclin-dependent kinase 1

CoIP:

Co-immunoprecipitation

DMEM:

Dulbecco's modified Eagle medium

DNA:

Deoxyribonucleic acid

dNTP:

Deoxynucleoside triphosphate

ECL:

Electrochemiluminescence

EDTA:

Ethylenediaminetetraacetic acid

FBS:

Fetal bovine serum

GFP:

Green fluorescent protein

HCMV:

Human cytomegalovirus

HEK:

Human Embryonic Kidney Cells

HEPES:

4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

HIV:

Human immunodeficiency virus

hpi:

Hours post-infection

HSV:

Herpes Simplex Virus

IFN:

Interferon

NF-kB:

Nuclear factor kappa light chain enhancer of activated B cells

MOI:

Multiplicity of infection

MS:

Mass spectrometry

PEI:

Polyethyleneimine

RNA:

Ribonucleic acid

SAMHD1:

SAM and HD Domain Containing Deoxynucleoside Triphosphate Triphosphohydrolase 1

SDS:

Sodium dodecyl sulfate

SV40:

Simian virus 40

TCID50 :

Tissue culture infectious dose

UL:

Unique long

WT:

Wild type

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Acknowledgements

We thank all the Herpesvirus and Molecular Virology Research Unit members for the helpful discussion and Roger Everett (University of Glasgow) for the pLKO-based lentiviral expression system.

Funding

This work was supported by the China National Natural Science Foundation (Grants 81371826 and 81572002 to ZQ. The funders had no role in study design, data collection, analysis, decision to publish, or manuscript preparation.

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Authors and Affiliations

  1. College of Vet Medicine, University of Sulaimani, Sulaymaniyah, Iraq

    Sirwan Sleman, Nahla Saeed & Dyary Othman

  2. Medical Laboratory Analysis, College of Health Science, Cihan University of Sulaimani, Sulaymaniyah, Iraq

    Hastyar Najmuldeen

  3. Unit of Herpesvirus and Molecular Virology, Key Laboratory of Molecular Virology and Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China

    Sirwan Sleman, Hongyun Hao & Zhikang Qian

  4. Biology Department, College of Sciences, University of Sulaimani, Sulaymaniyah, Iraq

    Paywast Jalal

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  1. Sirwan Sleman
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Sleman, S., Najmuldeen, H., Hao, H. et al. Human cytomegalovirus UL24 and UL43 products participate in SAMHD1 subcellular localization. VirusDis. 33, 383–396 (2022). https://doi.org/10.1007/s13337-022-00799-3

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