Viral infections enhance cancer risk and threaten host genome integrity. Although human cytomegalovirus (HCMV) proteins have been detected in a wide spectrum of human malignancies and HCMV infections have been implicated in tumorigenesis, the underlying mechanisms remain poorly understood. Here, we employed a range of experimental approaches, including single-molecule DNA fiber analysis, and showed that infection by any of the four commonly used HCMV strains: AD169, Towne, TB40E or VR1814 induced replication stress (RS), as documented by host-cell replication fork asymmetry and formation of 53BP1 foci. The HCMV-evoked RS triggered an ensuing host DNA damage response (DDR) and chromosomal instability in both permissive and non-permissive human cells, the latter being particularly relevant in the context of tumorigenesis, as such cells can survive and proliferate after HCMV infection. The viral major immediate early enhancer and promoter (MIEP) that controls expression of the viral genes IE72 (IE-1) and IE86 (IE-2), contains transcription-factor binding sites shared by promoters of cellular stress-response genes. We found that DNA damaging insults, including those relevant for cancer therapy, enhanced IE72/86 expression. Thus, MIEP has been evolutionary shaped to exploit host DDR. Ectopically expressed IE72 and IE86 also induced RS and increased genomic instability. Of clinical relevance, we show that undergoing standard-of-care genotoxic radio-chemotherapy in patients with HCMV-positive glioblastomas correlated with elevated HCMV protein markers after tumor recurrence. Collectively, these results are consistent with our proposed concept of HCMV hijacking transcription-factor binding sites shared with host stress-response genes. We present a model to explain the potential oncomodulatory effects of HCMV infections through enhanced replication stress, subverted DNA damage response and induced genomic instability.

Department of Biochemistry and Biophysics Stockholm University Stockholm Sweden

Department of Medical Biochemistry and Biophysics Division of Genome Biology Science for Life Laboratory Karolinska Institute Stockholm Sweden

Department of Medicine Unit of Microbial Pathogenesis Karolinska Institutet Stockholm Sweden

Department of Neurology Karolinska University Hospital Stockholm Sweden

Department of Neurosurgery Copenhagen University Hospital Rigshospitalet Copenhagen Denmark

Department of Neurosurgery Karolinska University Hospital Stockholm Sweden

Department of Pathology Copenhagen University Hospital Rigshospitalet Copenhagen Denmark

DNA Replication and Cancer Group Danish Cancer Society Research Center Copenhagen Denmark

Genome Integrity Danish Cancer Society Research Center Copenhagen Denmark

Genome Integrity Laboratory Institute of Molecular Genetics Prague Czech Republic

Institute of Biomedicine University of Turku Turku Finland

MediCity Research Laboratory University of Turku Turku Finland

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