Mitotic phosphorylation of the lamin B receptor by a serine/arginine kinase and p34(cdc2).
The lamin B receptor (LBR) is an integral protein of the inner nuclear membrane that is modified at interphase by a nuclear envelope-bound protein kinase. This enzyme (RS kinase) specifically phosphorylates arginine-serine dipeptide motifs located at the NH2-terminal domain of LBR and regulates its interactions with other nuclear envelope proteins. To compare the phosphorylation state of LBR during interphase and mitosis, we performed phosphopeptide mapping of in vitro and in vivo 32P-labeled LBR and analyzed a series of recombinant proteins and synthetic peptides. Our results show that LBR undergoes two types of mitotic phosphorylation mediated by the RS and the p34(cdc2) protein kinases, respectively. The RS kinase modifies similar sites at interphase and mitosis (i.e. Ser76, Ser78, Ser80, Ser82, Ser84), whereas p34(cdc2) mainly phosphorylates Ser71. These findings clarify the phosphorylation state of LBR during the cell cycle and provide new information for understanding the mechanisms responsible for nuclear envelope assembly and disassembly.
Nikolakaki
E
E
Laboratory of Biochemistry, School of Chemistry, The Aristotelian University of Thessaloniki, Thessaloniki 54 006, Greece.
Meier
J
J
Simos
G
G
Georgatos
S D
SD
Giannakouros
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001122:Arginine Kinase / Q000378:metabolism*; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002478:Cells, Cultured; D002645:Chickens; D008938:Mitosis; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000378:metabolism*; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D010748:Phosphopeptides / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D014422:Turkeys; D000097682:Lamin B Receptor
D001122:Arginine Kinase; D016203:CDC2 Protein Kinase; D010455:Peptides; D010748:Phosphopeptides; D017346:Protein Serine-Threonine Kinases; D018160:Receptors, Cytoplasmic and Nuclear; D000097682:Lamin B Receptor