title: Human Tra2 proteins jointly control a CHEK1 splicing switch among alternative and constitutive target exons.
abstract: Alternative splicing--the production of multiple messenger RNA isoforms from a single gene--is regulated in part by RNA binding proteins. While the RBPs transformer2 alpha (Tra2α) and Tra2β have both been implicated in the regulation of alternative splicing, their relative contributions to this process are not well understood. Here we find simultaneous--but not individual--depletion of Tra2α and Tra2β induces substantial shifts in splicing of endogenous Tra2β target exons, and that both constitutive and alternative target exons are under dual Tra2α-Tra2β control. Target exons are enriched in genes associated with chromosome biology including CHEK1, which encodes a key DNA damage response protein. Dual Tra2 protein depletion reduces expression of full-length CHK1 protein, results in the accumulation of the DNA damage marker γH2AX and decreased cell viability. We conclude Tra2 proteins jointly control constitutive and alternative splicing patterns via paralog compensation to control pathways essential to the maintenance of cell viability.
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Andrew
authors.initials: A
authors.lastname: Best
authors.affiliation: School of Computing Science, Claremont Tower, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
authors.forename: Katherine
authors.initials: K
authors.lastname: James
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Caroline
authors.initials: C
authors.lastname: Dalgliesh
authors.affiliation: Institute for Cellular Medicine, Newcastle University, Framlington Place, Newcastle NE2 4HH, UK.
authors.forename: Elaine
authors.initials: E
authors.lastname: Hong
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Mahsa
authors.initials: M
authors.lastname: Kheirolahi-Kouhestani
authors.affiliation: Faculty of Computer and Information Science, University of Ljubljana, Trzaska cesta 25, SI-1000, Ljubljana, Slovenia.
authors.forename: Tomaz
authors.initials: T
authors.lastname: Curk
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Yaobo
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Marina
authors.initials: M
authors.lastname: Danilenko
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Rafiq
authors.initials: R
authors.lastname: Hussain
authors.affiliation: 1] Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK [2] Institute of Cardiovascular Sciences, The University of Manchester, Manchester M13 9NT, UK.
authors.forename: Bernard
authors.initials: B
authors.lastname: Keavney
authors.affiliation: School of Computing Science, Claremont Tower, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
authors.forename: Anil
authors.initials: A
authors.lastname: Wipat
authors.affiliation: Department of Microbiology and Infectious Diseases, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada J1E 4K8.
authors.forename: Roscoe
authors.initials: R
authors.lastname: Klinck
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle NE2 4HH, UK.
authors.forename: Ian G
authors.initials: IG
authors.lastname: Cowell
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle NE2 4HH, UK.
authors.forename: Ka
authors.initials: K
authors.lastname: Cheong Lee
authors.affiliation: Institute for Cell and Molecular Biosciences, Newcastle University, Newcastle NE2 4HH, UK.
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Austin
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Julian P
authors.initials: JP
authors.lastname: Venables
authors.affiliation: Department of Microbiology and Infectious Diseases, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Québec, Canada J1E 4K8.
authors.forename: Benoit
authors.initials: B
authors.lastname: Chabot
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: Mauro
authors.initials: M
authors.lastname: Santibanez Koref
authors.affiliation: Institute for Cellular Medicine, Newcastle University, Framlington Place, Newcastle NE2 4HH, UK.
authors.forename: Alison
authors.initials: A
authors.lastname: Tyson-Capper
authors.affiliation: Institute of Genetic Medicine, Newcastle University, Central Parkway, Newcastle NE1 3BZ, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Elliott
mesh_terms: D017398:Alternative Splicing; D045744:Cell Line, Tumor; D000071877:Checkpoint Kinase 1; D005091:Exons; D006801:Humans; D061986:MCF-7 Cells; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D000068103:Serine-Arginine Splicing Factors
chemical_list: D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C000599452:TRA2A protein, human; C097083:TRA2B protein, human; D000068103:Serine-Arginine Splicing Factors; D011494:Protein Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1