title: Transcriptional silencing of long noncoding RNA GNG12-AS1 uncouples its transcriptional and product-related functions.
abstract: Long noncoding RNAs (lncRNAs) regulate gene expression via their RNA product or through transcriptional interference, yet a strategy to differentiate these two processes is lacking. To address this, we used multiple small interfering RNAs (siRNAs) to silence GNG12-AS1, a nuclear lncRNA transcribed in an antisense orientation to the tumour-suppressor DIRAS3. Here we show that while most siRNAs silence GNG12-AS1 post-transcriptionally, siRNA complementary to exon 1 of GNG12-AS1 suppresses its transcription by recruiting Argonaute 2 and inhibiting RNA polymerase II binding. Transcriptional, but not post-transcriptional, silencing of GNG12-AS1 causes concomitant upregulation of DIRAS3, indicating a function in transcriptional interference. This change in DIRAS3 expression is sufficient to impair cell cycle progression. In addition, the reduction in GNG12-AS1 transcripts alters MET signalling and cell migration, but these are independent of DIRAS3. Thus, differential siRNA targeting of a lncRNA allows dissection of the functions related to the process and products of its transcription.
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Lovorka
authors.initials: L
authors.lastname: Stojic
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Malwina
authors.initials: M
authors.lastname: Niemczyk
authors.affiliation: Biosearch Technologies Inc., 2199S. McDowell Boulevard, Petaluma, California 94954, USA.
authors.forename: Arturo
authors.initials: A
authors.lastname: Orjalo
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Yoko
authors.initials: Y
authors.lastname: Ito
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Anna Elisabeth Maria
authors.initials: AE
authors.lastname: Ruijter
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Santiago
authors.initials: S
authors.lastname: Uribe-Lewis
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Nimesh
authors.initials: N
authors.lastname: Joseph
authors.affiliation: Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, UK.
authors.forename: Stephen
authors.initials: S
authors.lastname: Weston
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Suraj
authors.initials: S
authors.lastname: Menon
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Duncan T
authors.identifier: http://orcid.org/0000-0001-6201-5599
authors.initials: DT
authors.lastname: Odom
authors.affiliation: Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
authors.forename: John
authors.initials: J
authors.lastname: Rinn
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.
authors.forename: Fanni
authors.initials: F
authors.lastname: Gergely
authors.affiliation: Cancer Research UK Cambridge Institute, University of Cambridge, Li Ka Shing Centre, Robinson Way, Cambridge CB2 0RE, UK.|Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath BA2 7AY, UK.
authors.forename: Adele
authors.initials: A
authors.lastname: Murrell
mesh_terms: D060565:Argonaute Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D044388:GTP-Binding Protein gamma Subunits / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D020033:Protein Isoforms; D034622:RNA Interference; D012320:RNA Polymerase III / Q000235:genetics / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000235:genetics / Q000378:metabolism*; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C466422:AGO2 protein, human; D060565:Argonaute Proteins; C117386:DIRAS3 protein, human; D044388:GTP-Binding Protein gamma Subunits; C548441:Gng12 protein, mouse; D020033:Protein Isoforms; D062085:RNA, Long Noncoding; C000608316:long noncoding RNA GNG12-AS1, human; D012320:RNA Polymerase III; D020741:rho GTP-Binding Proteins
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