title: Artemis C-terminal region facilitates V(D)J recombination through its interactions with DNA Ligase IV and DNA-PKcs.
abstract: Artemis is an endonuclease that opens coding hairpin ends during V(D)J recombination and has critical roles in postirradiation cell survival. A direct role for the C-terminal region of Artemis in V(D)J recombination has not been defined, despite the presence of immunodeficiency and lymphoma development in patients with deletions in this region. Here, we report that the Artemis C-terminal region directly interacts with the DNA-binding domain of Ligase IV, a DNA Ligase which plays essential roles in DNA repair and V(D)J recombination. The Artemis-Ligase IV interaction is specific and occurs independently of the presence of DNA and DNA-protein kinase catalytic subunit (DNA-PKcs), another protein known to interact with the Artemis C-terminal region. Point mutations in Artemis that disrupt its interaction with Ligase IV or DNA-PKcs reduce V(D)J recombination, and Artemis mutations that affect interactions with Ligase IV and DNA-PKcs show additive detrimental effects on coding joint formation. Signal joint formation remains unaffected. Our data reveal that the C-terminal region of Artemis influences V(D)J recombination through its interaction with both Ligase IV and DNA-PKcs.
authors.affiliation: Immunology Institute, Mount Sinai School of Medicine, New York, NY 10029, USA.
authors.forename: Shruti
authors.initials: S
authors.lastname: Malu
authors.forename: Pablo
authors.initials: P
authors.lastname: De Ioannes
authors.forename: Mikhail
authors.initials: M
authors.lastname: Kozlov
authors.forename: Marsha
authors.initials: M
authors.lastname: Greene
authors.forename: Dailia
authors.initials: D
authors.lastname: Francis
authors.forename: Mary
authors.initials: M
authors.lastname: Hanna
authors.forename: Jesse
authors.initials: J
authors.lastname: Pena
authors.forename: Carlos R
authors.initials: CR
authors.lastname: Escalante
authors.forename: Aya
authors.initials: A
authors.lastname: Kurosawa
authors.forename: Hediye
authors.initials: H
authors.lastname: Erdjument-Bromage
authors.forename: Paul
authors.initials: P
authors.lastname: Tempst
authors.forename: Noritaka
authors.initials: N
authors.lastname: Adachi
authors.forename: Paolo
authors.initials: P
authors.lastname: Vezzoni
authors.forename: Anna
authors.initials: A
authors.lastname: Villa
authors.forename: Aneel K
authors.initials: AK
authors.lastname: Aggarwal
authors.forename: Patricia
authors.initials: P
authors.lastname: Cortes
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D004260:DNA Repair / Q000235:genetics; D051747:DNA-Activated Protein Kinase / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004720:Endonucleases; D005454:Fluorescence Polarization; D005822:Genetic Vectors / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D017354:Point Mutation / Q000235:genetics; D017422:Sequence Analysis, DNA; D014162:Transfection; D060152:V(D)J Recombination / Q000502:physiology*
chemical_list: D017931:DNA Primers; D004268:DNA-Binding Proteins; C000607893:LIG4 protein, human; D009687:Nuclear Proteins; C101467:XRCC4 protein, human; D051747:DNA-Activated Protein Kinase; C497263:PRKDC protein, human; C426344:DCLRE1C protein, human; D004720:Endonucleases; D011088:DNA Ligases; D000072481:DNA Ligase ATP