Multisite phosphorylation of nuclear interaction partner of ALK (NIPA) at G2/M involves cyclin B1/Cdk1.
Nuclear interaction partner of ALK (NIPA) is an F-box-containing protein that defines a nuclear skp1 cullin F-box (SCF)-type ubiquitin E3 ligase (SCFNIPA) implicated in the regulation of mitotic entry. The SCFNIPA complex targets nuclear cyclin B1 for ubiquitination in interphase, whereas phosphorylation of NIPA in late G2 phase and mitosis inactivates the complex to allow for accumulation of cyclin B1. Here, we identify the region of NIPA that mediates binding to its substrate cyclin B1. In addition to the recently described serine residue 354, we specify 2 new residues, Ser-359 and Ser-395, implicated in the phosphorylation process at G2/M within this region. Moreover, we found cyclin B1/Cdk1 to phosphorylate NIPA at Ser-395 in mitosis. Mutation of both Ser-359 and Ser-395 impaired effective inactivation of the SCFNIPA complex, resulting in reduced levels of mitotic cyclin B1. These data are compatible with a process of sequential NIPA phosphorylation where cyclin B1/Cdk1 amplifies phosphorylation of NIPA once an initial phosphorylation event has dissociated the SCFNIPA complex. Thus, cyclin B1/Cdk1 may contribute to the regulation of its own abundance in early mitosis.
Bassermann
Florian
F
Department of Internal Medicine III, Technical University of Munich, 81675 Munich, Germany, and Department of Pathology, St. Jude Chidren's Research Hospital, Memphis, TN 38105, USA. florian.bassermann@med.nyu.edu
von Klitzing
Christine
C
Illert
Anna Lena
AL
Münch
Silvia
S
Morris
Stephan W
SW
Pagano
Michele
M
Peschel
Christian
C
Duyster
Justus
J
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D019556:COS Cells; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D002522:Chlorocebus aethiops; D019926:Cyclin B / Q000378:metabolism*; D056744:Cyclin B1; D016195:G2 Phase / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D008938:Mitosis / Q000502:physiology*; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D044843:SKP Cullin F-Box Protein Ligases / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; C534699:CCNB1 protein, human; D018797:Cell Cycle Proteins; D019926:Cyclin B; D056744:Cyclin B1; D009687:Nuclear Proteins; D025801:Ubiquitin; C477288:ZC3HC1 protein, human; D044843:SKP Cullin F-Box Protein Ligases; D016203:CDC2 Protein Kinase