Cutoff Suppresses RNA Polymerase II Termination to Ensure Expression of piRNA Precursors.
Small non-coding RNAs called piRNAs serve as guides for an adaptable immune system that represses transposable elements in germ cells of Metazoa. In Drosophila the RDC complex, composed of Rhino, Deadlock and Cutoff (Cuff) bind chromatin of dual-strand piRNA clusters, special genomic regions, which encode piRNA precursors. The RDC complex is required for transcription of piRNA precursors, though the mechanism by which it licenses transcription remained unknown. Here, we show that Cuff prevents premature termination of RNA polymerase II. Cuff prevents cleavage of nascent RNA at poly(A) sites by interfering with recruitment of the cleavage and polyadenylation specificity factor (CPSF) complex. Cuff also protects processed transcripts from degradation by the exonuclease Rat1. Our work reveals a conceptually different mechanism of transcriptional enhancement. In contrast to other factors that regulate termination by binding to specific signals on nascent RNA, the RDC complex inhibits termination in a chromatin-dependent and sequence-independent manner.
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Yung-Chia Ariel
YA
Chen
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.|Biochemistry Center Regensburg, Laboratory for RNA Biology, University of Regensburg, 93053 Regensburg, Germany.
Evelyn
E
Stuwe
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Yicheng
Y
Luo
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Maria
M
Ninova
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Adrien
A
Le Thomas
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Ekaterina
E
Rozhavskaya
Memorial Sloan-Kettering Cancer Center, Structural Biology Program, 1275 York Avenue, New York, NY, 10021 USA.
Sisi
S
Li
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Sivani
S
Vempati
Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8.
John D
JD
Laver
Memorial Sloan-Kettering Cancer Center, Structural Biology Program, 1275 York Avenue, New York, NY, 10021 USA.
Dinshaw J
DJ
Patel
Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8.|Department of Biochemistry, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8.
Craig A
CA
Smibert
Department of Molecular Genetics, University of Toronto, 1 King's College Circle, Toronto, Ontario, Canada M5S 1A8.
Howard D
HD
Lipshitz
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Katalin Fejes
KF
Toth
California Institute of Technology, Division of Biology, 147-75, 1200 E. California Blvd., Pasadena, CA 91125, USA.
Alexei A
AA
Aravin
D000241:Adenosine / Q000378:metabolism; D000818:Animals; D030801:Animals, Genetically Modified; D001665:Binding Sites; D002843:Chromatin / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D039223:Cleavage And Polyadenylation Specificity Factor / Q000378:metabolism; D019295:Computational Biology; D030541:Databases, Genetic; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D005095:Exoribonucleases / Q000378:metabolism; D017930:Genes, Reporter; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D046912:Multiprotein Complexes; D011108:Polymers / Q000378:metabolism; D011485:Protein Binding; D012319:RNA Polymerase II / Q000378:metabolism*; D020871:RNA Stability; D034741:RNA, Small Interfering / Q000096:biosynthesis* / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D061806:Transcription Termination, Genetic; D014158:Transcription, Genetic
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D039223:Cleavage And Polyadenylation Specificity Factor; C000590395:Cuff protein, Drosophila; D029721:Drosophila Proteins; D008869:Microtubule-Associated Proteins; D046912:Multiprotein Complexes; D011108:Polymers; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; C583035:del protein, Drosophila; C449641:rhi protein, Drosophila; C069424:polyadenosine; D012319:RNA Polymerase II; D005095:Exoribonucleases; D000241:Adenosine