title: Modulation of homology-directed repair in T98G glioblastoma cells due to interactions between wildtype p53, Rad51 and HCMV IE1-72.
abstract: Human cytomegalovirus (HCMV) is a ubiquitous pathogen capable of causing life threatening consequences in neonates and immune-compromised individuals. HCMV inflicts site-specific double strand breaks (DSBs) in the cellular genome. DNA damage infliction raises the corollary question of virus modulation of DNA repair. We recently reported HDR was stimulated in wt human foreskin fibroblasts (HFFs) during fully permissive infection or expression of the HCMV protein IE1-72 (IE72). These studies have been extended into semi-permissive T98G glioblastoma cells. T98Gs encode a mutant p53, which may contribute to their high baseline rate of HDR. We fully expected HCMV infection to increase HDR in T98Gs, similar to its effects in HFFs. Surprisingly in T98Gs HCMV infection, or sole expression of IE72, decreased HDR by two-fold. Transient expression of wt p53 in T98Gs also reduced HDR by two-fold. Dual transient expression of wt p53 and IE72 restored high baseline HDR levels. GST pulldown experiments revealed that both IE72 and wt p53 bound the important HDR protein, Rad51. We conclude that the expression of certain HCMV proteins can modulate HDR in an infected cell, dependent upon p53 status. We propose a model of the protein interactions explaining this behavior.
authors.affiliation: Tumorvirologie (F010), Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, 69120, Heidelberg, Germany. amitkulkar@gmail.com.
authors.forename: Amit S
authors.initials: AS
authors.lastname: Kulkarni
authors.affiliation: Department of Biological Sciences and the Center for Reproductive Biology, University of Idaho, 875 Perimeter Drive, Mailstop 3051, Moscow, ID 83844, USA. lfort@uidaho.edu.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Fortunato
mesh_terms: D045744:Cell Line, Tumor; D003587:Cytomegalovirus / Q000254:growth & development*; D004260:DNA Repair; D054884:Host-Pathogen Interactions; D006801:Humans; D017874:Immediate-Early Proteins / Q000378:metabolism*; D011485:Protein Binding; D051135:Rad51 Recombinase / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: C088645:IE1 protein, cytomegalovirus; D017874:Immediate-Early Proteins; D016159:Tumor Suppressor Protein p53; C495153:RAD51 protein, human; D051135:Rad51 Recombinase