title: RNA polymerase II synthesizes antisense RNA in Plasmodium falciparum.
abstract: The recent identification of antisense RNA in the transcriptomes of many eukaryotes has generated enormous interest. The presence of antisense RNA in Plasmodium falciparum, the causative agent of severe malaria, remains controversial. Elucidation of the mechanism of antisense RNA in P. falciparum synthesis is critical in order to demonstrate the origin and function of these transcripts. Therefore, a systematic analysis of antisense and sense RNA synthesis was performed using direct labeling experiments. Nuclear run on experiments with single-stranded DNA probes demonstrated that antisense RNA is synthesized in the nucleus at several genomic loci. Antisense RNA synthesis is sensitive to the potent RNA polymerase II inhibitor alpha-amanitin. Antisense and sense transcription was also detected in nuclei isolated from synchronized parasites, suggesting concurrent synthesis. In summary, our experiments directly demonstrate that antisense RNA synthesis is a common transcriptional phenomenon in P. falciparum, and is catalyzed by RNA polymerase II.
authors.affiliation: Harvard School of Public Health, Department of Immunology and Infectious Diseases, 665 Huntington Avenue, Building I, Room 705, Boston, MA 02115, USA.
authors.forename: Kevin T
authors.initials: KT
authors.lastname: Militello
authors.forename: Vishal
authors.initials: V
authors.lastname: Patel
authors.forename: Anne-Danielle
authors.initials: AD
authors.lastname: Chessler
authors.forename: Julia K
authors.initials: JK
authors.lastname: Fisher
authors.forename: Jacob M
authors.initials: JM
authors.lastname: Kasper
authors.forename: Anusha
authors.initials: A
authors.lastname: Gunasekera
authors.forename: Dyann F
authors.initials: DF
authors.lastname: Wirth
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D004277:DNA, Single-Stranded; D010963:Plasmodium falciparum / Q000235:genetics* / Q000378:metabolism; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism*; D016372:RNA, Antisense / Q000096:biosynthesis* / Q000235:genetics; D016053:RNA, Protozoan / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: D004277:DNA, Single-Stranded; D016372:RNA, Antisense; D016053:RNA, Protozoan; D012319:RNA Polymerase II