M-phase-specific phosphorylation and structural rearrangement of the cytoplasmic cross-linking protein plectin involve p34cdc2 kinase.
Plectin, a widespread and abundant cytoskeletal cross-linking protein, serves as a target for protein kinases throughout the cell cycle, without any significant variation in overall phosphorylation level. One of the various phosphorylation sites of the molecule was found to be phosphorylated preferentially during mitosis. By in vivo phosphorylation of ectopically expressed plectin domains in stably transfected Chinese hamster ovary cells, this site was mapped to the C-terminal repeat 6 domain of the polypeptide. The same site has been identified as an in vitro target for p34cdc2 kinase. Mitosis-specific phosphorylation of plectin was accompanied by a rearrangement of plectin structures, changing from a filamentous, largely vimentin-associated state in interphase to a diffuse vimentin-independent distribution in mitosis as visualized by immunofluorescence microscopy. Subcellular fractionation studies showed that in interphase cells up to 80% of cellular plectin was found associated with an insoluble cell fraction mostly consisting of intermediate filaments, while during mitosis the majority of plectin (> 75%) became soluble. Furthermore, phosphorylation of purified plectin by p34cdc2 kinase decreased plectin's ability to interact with preassembled vimentin filaments in vitro. Together, our data suggest that a mitosis-specific phosphorylation involving p34cdc2 kinase regulates plectin's cross-linking activities and association with intermediate filaments during the cell cycle.
Foisner
R
R
Institute of Biochemistry and Molecular Cell Biology, University of Vienna, Austria.
Malecz
N
N
Dressel
N
N
Stadler
C
C
Wiche
G
G
D000595:Amino Acid Sequence; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D016466:CHO Cells; D002453:Cell Cycle / Q000502:physiology; D006224:Cricetinae; D003593:Cytoplasm; D007381:Intermediate Filament Proteins / Q000737:chemistry / Q000378:metabolism*; D007382:Intermediate Filaments / Q000378:metabolism; D008938:Mitosis; D008969:Molecular Sequence Data; D010766:Phosphorylation; D051190:Plectin
D007381:Intermediate Filament Proteins; D051190:Plectin; D016203:CDC2 Protein Kinase