title: A Bioorthogonal Chemical Reporter of Viral Infection.
abstract: Pathogen-selective labeling was achieved by using the novel gemcitabine metabolite analogue 2'-deoxy-2',2'-difluoro-5-ethynyluridine (dF-EdU) and click chemistry. Cells infected with Herpes Simplex Virus-1 (HSV-1), but not uninfected cells, exhibit nuclear staining upon the addition of dF-EdU and a fluorescent azide. The incorporation of the dF-EdU into DNA depends on its phosphorylation by a herpes virus thymidine kinase (TK). Crystallographic analyses revealed how dF-EdU is well accommodated in the active site of HSV-1 TK, but steric clashes prevent dF-EdU from binding human TK. These results provide the first example of pathogen-enzyme-dependent incorporation and labeling of bioorthogonal functional groups in human cells.
authors.affiliation: Department of Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich (Switzerland) http://www.bioorganic-chemistry.com.
authors.forename: Anne B
authors.initials: AB
authors.lastname: Neef
authors.affiliation: Pharmaceutical Biochemistry, University of Geneva (Switzerland).
authors.forename: Lucile
authors.initials: L
authors.lastname: Pernot
authors.affiliation: Department of Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich (Switzerland) http://www.bioorganic-chemistry.com.
authors.forename: Verena N
authors.initials: VN
authors.lastname: Schreier
authors.affiliation: Pharmaceutical Biochemistry, University of Geneva (Switzerland).
authors.forename: Leonardo
authors.initials: L
authors.lastname: Scapozza
authors.affiliation: Department of Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich (Switzerland) http://www.bioorganic-chemistry.com. nathan.luedtke@chem.uzh.ch.
authors.forename: Nathan W
authors.initials: NW
authors.lastname: Luedtke
mesh_terms: D000818:Animals; D001386:Azides / Q000737:chemistry* / Q000378:metabolism; D020134:Catalytic Domain; D002522:Chlorocebus aethiops; D057930:Click Chemistry; D005456:Fluorescent Dyes / Q000737:chemistry* / Q000378:metabolism; D054879:Halogenation; D006367:HeLa Cells; D006561:Herpes Simplex / Q000175:diagnosis* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000201:enzymology / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D013194:Staining and Labeling; D013937:Thymidine Kinase / Q000032:analysis / Q000378:metabolism; D014529:Uridine / Q000031:analogs & derivatives* / Q000378:metabolism; D014709:Vero Cells
chemical_list: D001386:Azides; D005456:Fluorescent Dyes; D013937:Thymidine Kinase; D014529:Uridine
keywords: bioorthogonal chemistry; chemical reporters; click chemistry; nucleosides; viruses