Nucleolin, defective for MPF phosphorylation, localizes normally during mitosis and nucleologenesis.
To determine what effect maturation promoting factor (MPF, p34(cdc2) kinase/cyclin B) phosphorylation has on nucleolin's distribution during mitotic nucleolar disassembly and reassembly, we altered Chinese hamster ovary (CHO) nucleolin (the N protein) such that it cannot be phosphorylated by p34(cdc2). As expected, the transiently expressed epitope-tagged N protein showed no apparent defect in nucleolar localization in interphase CHO cells, even after hypotonic shock and recovery to quickly disassemble and then reassemble interphase nucleoli. In mitotic CHO cells, the N protein localized to the perichromosomal sheath and the cytoplasm, as is typical for nucleolin. Similar to epitope-tagged wild-type nucleolin, the N protein also maintained its association with persistent nucleoli characteristic of mitotic Chinese hamster lung (Dede) cells. In synchronized HeLa cells, the N protein again localized to the perichromosomal sheath and the cytoplasm as nucleoli disassembled during prophase. In HeLa cell telophase, the N protein localized normally to nucleolus-derived foci within the cytoplasm and prenucleolar bodies within reforming nuclei. The observations indicate that MPF phosphorylation is not essential for nucleolin's localizations to the perichromosomal sheath and the cytoplasm during prophase and metaphase, and that functional MPF phosphorylation sites are not essential for nucleolin's localizations during nucleologenesis.
Zhu
Y
Y
Department of Biological Sciences, Louisiana State University, Baton Rouge 70803-1806, USA.
Lu
D
D
DiMario
P
P
D000595:Amino Acid Sequence; D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D006224:Cricetinae; D019926:Cyclin B / Q000378:metabolism; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D016200:Maturation-Promoting Factor / Q000378:metabolism*; D008938:Mitosis; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000097796:Nucleolin
D016203:CDC2 Protein Kinase; D019926:Cyclin B; D016200:Maturation-Promoting Factor; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D000097796:Nucleolin