title: Glucosylated hydroxymethyluracil, DNA base J, prevents transcriptional readthrough in Leishmania.
abstract: Some Ts in nuclear DNA of trypanosomes and Leishmania are hydroxylated and glucosylated to yield base J (β-D-glucosyl-hydroxymethyluracil). In Leishmania, about 99% of J is located in telomeric repeats. We show here that most of the remaining J is located at chromosome-internal RNA polymerase II termination sites. This internal J and telomeric J can be reduced by a knockout of J-binding protein 2 (JBP2), an enzyme involved in the first step of J biosynthesis. J levels are further reduced by growing Leishmania JBP2 knockout cells in BrdU-containing medium, resulting in cell death. The loss of internal J in JBP2 knockout cells is accompanied by massive readthrough at RNA polymerase II termination sites. The readthrough varies between transcription units but may extend over 100 kb. We conclude that J is required for proper transcription termination and infer that the absence of internal J kills Leishmania by massive readthrough of transcriptional stops.
authors.affiliation: Division of Molecular Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: Henri G A M
authors.initials: HG
authors.lastname: van Luenen
authors.forename: Carol
authors.initials: C
authors.lastname: Farris
authors.forename: Sabrina
authors.initials: S
authors.lastname: Jan
authors.forename: Paul-Andre
authors.initials: PA
authors.lastname: Genest
authors.forename: Pankaj
authors.initials: P
authors.lastname: Tripathi
authors.forename: Arno
authors.initials: A
authors.lastname: Velds
authors.forename: Ron M
authors.initials: RM
authors.lastname: Kerkhoven
authors.forename: Marja
authors.initials: M
authors.lastname: Nieuwland
authors.forename: Andrew
authors.initials: A
authors.lastname: Haydock
authors.forename: Gowthaman
authors.initials: G
authors.lastname: Ramasamy
authors.forename: Saara
authors.initials: S
authors.lastname: Vainio
authors.forename: Tatjana
authors.initials: T
authors.lastname: Heidebrecht
authors.forename: Anastassis
authors.initials: A
authors.lastname: Perrakis
authors.forename: Ludo
authors.initials: L
authors.lastname: Pagie
authors.forename: Bas
authors.initials: B
authors.lastname: van Steensel
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Myler
authors.forename: Piet
authors.initials: P
authors.lastname: Borst
mesh_terms: D055786:Gene Knockout Techniques; D005960:Glucosides / Q000378:metabolism*; D007891:Leishmania / Q000235:genetics* / Q000378:metabolism*; D012319:RNA Polymerase II / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000378:metabolism; D014158:Transcription, Genetic; D014498:Uracil / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D005960:Glucosides; D012330:RNA, Double-Stranded; C084880:5-((glucopyranosyloxy)methyl)uracil; D014498:Uracil; D012319:RNA Polymerase II