title: Identification of novel DNA-damage tolerance genes reveals regulation of translesion DNA synthesis by nucleophosmin.
abstract: Cells cope with replication-blocking lesions via translesion DNA synthesis (TLS). TLS is carried out by low-fidelity DNA polymerases that replicate across lesions, thereby preventing genome instability at the cost of increased point mutations. Here we perform a two-stage siRNA-based functional screen for mammalian TLS genes and identify 17 validated TLS genes. One of the genes, NPM1, is frequently mutated in acute myeloid leukaemia (AML). We show that NPM1 (nucleophosmin) regulates TLS via interaction with the catalytic core of DNA polymerase-η (polη), and that NPM1 deficiency causes a TLS defect due to proteasomal degradation of polη. Moreover, the prevalent NPM1c+ mutation that causes NPM1 mislocalization in ~30% of AML patients results in excessive degradation of polη. These results establish the role of NPM1 as a key TLS regulator, and suggest a mechanism for the better prognosis of AML patients carrying mutations in NPM1.
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Omer
authors.initials: O
authors.lastname: Ziv
authors.affiliation: Department of Physics of Complex Systems, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Amit
authors.initials: A
authors.lastname: Zeisel
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Nataly
authors.initials: N
authors.lastname: Mirlas-Neisberg
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Umakanta
authors.initials: U
authors.lastname: Swain
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Reinat
authors.initials: R
authors.lastname: Nevo
authors.affiliation: Department of Biological Regulation, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Nir
authors.initials: N
authors.lastname: Ben-Chetrit
authors.affiliation: Institute of Hematology, University of Perugia, Perugia 06100, Italy.
authors.forename: Maria Paola
authors.initials: MP
authors.lastname: Martelli
authors.affiliation: Institute of Hematology, University of Perugia, Perugia 06100, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Rossi
authors.affiliation: Department of Chemistry, Ludwig Maximilians University Munich, Munich 81377, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schiesser
authors.affiliation: Department of Pharmacology, University of Michigan, Ann Arbor, Michigan 48109, USA.
authors.forename: Christine E
authors.initials: CE
authors.lastname: Canman
authors.affiliation: Department of Chemistry, Ludwig Maximilians University Munich, Munich 81377, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Carell
authors.affiliation: Department of Chemistry, New York University, New York, New York 10003, USA.
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Geacintov
authors.affiliation: Institute of Hematology, University of Perugia, Perugia 06100, Italy.
authors.forename: Brunangelo
authors.initials: B
authors.lastname: Falini
authors.affiliation: Department of Physics of Complex Systems, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Eytan
authors.initials: E
authors.lastname: Domany
authors.affiliation: Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Zvi
authors.initials: Z
authors.lastname: Livneh
mesh_terms: D002460:Cell Line; D004249:DNA Damage / Q000528:radiation effects; D004260:DNA Repair; D004261:DNA Replication / Q000528:radiation effects; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D000090243:Nucleophosmin; D011485:Protein Binding; D014466:Ultraviolet Rays; D000099270:Y-Family DNA Polymerases
chemical_list: D004259:DNA-Directed DNA Polymerase; D009687:Nuclear Proteins; D000090243:Nucleophosmin; C000716967:NPM1 protein, human; C112338:Rad30 protein; D000099270:Y-Family DNA Polymerases