title: Regulation of alternative polyadenylation by Nkx2-5 and Xrn2 during mouse heart development.
abstract: Transcription factors organize gene expression profiles by regulating promoter activity. However, the role of transcription factors after transcription initiation is poorly understood. Here, we show that the homeoprotein Nkx2-5 and the 5'-3' exonuclease Xrn2 are involved in the regulation of alternative polyadenylation (APA) during mouse heart development. Nkx2-5 occupied not only the transcription start sites (TSSs) but also the downstream regions of genes, serving to connect these regions in primary embryonic cardiomyocytes (eCMs). Nkx2-5 deficiency affected Xrn2 binding to target loci and resulted in increases in RNA polymerase II (RNAPII) occupancy and in the expression of mRNAs with long 3'untranslated regions (3' UTRs) from genes related to heart development. siRNA-mediated suppression of Nkx2-5 and Xrn2 led to heart looping anomaly. Moreover, Nkx2-5 genetically interacts with Xrn2 because Nkx2-5(+/-)Xrn2(+/-), but neither Nkx2-5(+/-)nor Xrn2(+/-), newborns exhibited a defect in ventricular septum formation, suggesting that the association between Nkx2-5 and Xrn2 is essential for heart development. Our results indicate that Nkx2-5 regulates not only the initiation but also the usage of poly(A) sites during heart development. Our findings suggest that tissue-specific transcription factors is involved in the regulation of APA.
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Keisuke
authors.identifier: 0000-0001-9680-2646
authors.initials: K
authors.lastname: Nimura
authors.affiliation: Department of Nephrology, Kyoto University Graduate School of Medicine, Kyoto, Japan.
authors.forename: Masamichi
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Takeichi
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Kotaro
authors.initials: K
authors.lastname: Saga
authors.affiliation: Developmental Genetics Group, Graduate School of Frontier Biosciences, Osaka University, Suita, Japan.
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Takaoka
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.|Department of Urology, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Norihiko
authors.initials: N
authors.lastname: Kawamura
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Nitta
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Nagano
authors.affiliation: Center for Medical Research and Education, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Saki
authors.initials: S
authors.lastname: Ishino
authors.affiliation: Center for Medical Research and Education, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: Department of Biology and Biochemistry, University of Houston, Houston, Unites States.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Schwartz
authors.affiliation: Genome Science Division, Research Center for Advanced Science and Technology (RCAST), The University of Tokyo, Tokyo, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Aburatani
authors.affiliation: Division of Gene Therapy Science, Osaka University Graduate School of Medicine, Suita, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Kaneda
mesh_terms: D000818:Animals; D005095:Exoribonucleases / Q000235:genetics / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D006321:Heart / Q000196:embryology*; D000072576:Homeobox Protein Nkx-2.5 / Q000235:genetics / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D026723:Polyadenylation
chemical_list: D000072576:Homeobox Protein Nkx-2.5; C484469:Nkx2-5 protein, mouse; D005095:Exoribonucleases; C092452:Xrn2 protein, mouse
keywords: chromosomes; development; gene expression; genes; mouse; transcription factors