Cyclin F regulates the nuclear localization of cyclin B1 through a cyclin–cyclin interaction

The key regulator of G2–M transition of the cell cycle is M‐phase promoting factor (MPF), a complex composed of cdc2 and a B‐type cyclin. Cyclin B1 nuclear localization involves phosphorylation within a region called the cytoplasmic retention signal, which also contains a nuclear export signal. The mechanism of MPF nuclear localization remains unclear since it contains no functional nuclear localization signal (NLS). We exploited the yeast two‐hybrid screen to find protein(s) potentially mediating localization of cyclin B1 and identified a novel interaction between cyclin B1 and cyclin F. We found that cdc2, cyclin B1 and cyclin F form a complex that exhibits histone H1 kinase activity. Cyclin B1 and cyclin F also colocalize through immunofluorescence studies. Additionally, deletion analysis revealed that each putative NLS of cyclin F is functional. Taken together, the data suggest that the NLS regions of cyclin F regulate cyclin B1 localization to the nucleus. The interaction between cyclin B1 and cyclin F represents the first example of direct cyclin–cyclin binding, and elucidates a novel mechanism that regulates MPF localization and function.

[1]  E. Nigg,et al.  Nuclear localization of vertebrate cyclin A correlates with its ability to form complexes with cdk catalytic subunits. , 1993, Journal of cell science.

[2]  A. Murray,et al.  Cyclin is degraded by the ubiquitin pathway , 1991, Nature.

[3]  Eric T. Rosenthal,et al.  Cyclin: A protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division , 1983, Cell.

[4]  T. Hunt,et al.  Identification of the domains in cyclin A required for binding to, and activation of, p34cdc2 and p32cdk2 protein kinase subunits. , 1992, Molecular biology of the cell.

[5]  L. Meijer,et al.  Cyclin is a component of the sea urchin egg M‐phase specific histone H1 kinase. , 1989, The EMBO journal.

[6]  F. Cross,et al.  An N-terminal peptide from p60src can direct myristylation and plasma membrane localization when fused to heterologous proteins , 1985, Nature.

[7]  T. Hunter,et al.  Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport , 1991, The Journal of cell biology.

[8]  T. Hunt,et al.  Cyclins and their partners: from a simple idea to complicated reality. , 1991, Seminars in cell biology.

[9]  J. Pines,et al.  MPF localization is controlled by nuclear export , 1998, The EMBO journal.

[10]  Stephen J. Elledge,et al.  SKP1 Connects Cell Cycle Regulators to the Ubiquitin Proteolysis Machinery through a Novel Motif, the F-Box , 1996, Cell.

[11]  D. Beach,et al.  Activation of M-phase-specific histone H1 kinase by modification of the phosphorylation of its p34cdc2 and cyclin components. , 1990, Genes & development.

[12]  J. Ruderman,et al.  The role of cyclin B in meiosis I , 1989, The Journal of cell biology.

[13]  Jonathan A. Cooper,et al.  Mammalian Ras interacts directly with the serine/threonine kinase raf , 1993, Cell.

[14]  Paul Russell,et al.  Negative regulation of mitosis by wee1 +, a gene encoding a protein kinase homolog , 1987, Cell.

[15]  G. Blobel,et al.  Mammalian karyopherin alpha 1 beta and alpha 2 beta heterodimers: alpha 1 or alpha 2 subunit binds nuclear localization signal and beta subunit interacts with peptide repeat-containing nucleoporins. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[16]  David Beach,et al.  cdc2 protein kinase is complexed with both cyclin A and B: Evidence for proteolytic inactivation of MPF , 1989, Cell.

[17]  Marc W. Kirschner,et al.  cdc25 is a specific tyrosine phosphatase that directly activates p34cdc2 , 1991, Cell.

[18]  S. Elledge,et al.  Human cyclin F. , 1994, The EMBO journal.

[19]  T. Coleman,et al.  Myt1: A Membrane-Associated Inhibitory Kinase That Phosphorylates Cdc2 on Both Threonine-14 and Tyrosine-15 , 1995, Science.

[20]  J. Maller,et al.  Maternal Xenopus Cdk2-Cyclin E Complexes Function during Meiotic and Early Embryonic Cell Cycles That Lack a G1 Phase (*) , 1995, The Journal of Biological Chemistry.

[21]  T. Hunt,et al.  The cdc2‐related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. , 1993, The EMBO journal.

[22]  H. Lodish,et al.  Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface , 1986, Cell.

[23]  A. N. Meyer,et al.  Nuclear localization of cyclin B1 mediates its biological activity and is regulated by phosphorylation. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[24]  H. Piwnica-Worms,et al.  p107wee1 is a dual-specificity kinase that phosphorylates p34cdc2 on tyrosine 15. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[25]  F. Cross,et al.  A short sequence in the p60src N terminus is required for p60src myristylation and membrane association and for cell transformation , 1984, Molecular and cellular biology.

[26]  C. Alfa,et al.  Conserved structural motifs in cyclins identified by sequence analysis. , 1991, Journal of cell science.

[27]  H. Okayama,et al.  High-efficiency transformation of mammalian cells by plasmid DNA. , 1987, Molecular and cellular biology.

[28]  E. Nigg,et al.  Differential phosphorylation of vertebrate p34cdc2 kinase at the G1/S and G2/M transitions of the cell cycle: identification of major phosphorylation sites. , 1991, The EMBO journal.

[29]  P. Nurse.  Universal control mechanism regulating onset of M-phase , 1990, Nature.

[30]  K. Yamashita,et al.  The cdc2‐related protein p40MO15 is the catalytic subunit of a protein kinase that can activate p33cdk2 and p34cdc2. , 1993 .

[31]  T. Boulikas,et al.  Nuclear import of protein kinases and cyclins , 1996, Journal of cellular biochemistry.

[32]  R. Honda,et al.  Dephosphorylation of human p34 cdc2 kinase on both Thr‐14 and Tyr‐15 by human cdc25B phosphatase , 1993, FEBS letters.

[33]  Mike Tyers,et al.  F-Box Proteins Are Receptors that Recruit Phosphorylated Substrates to the SCF Ubiquitin-Ligase Complex , 1997, Cell.

[34]  Kathleen L. Gould,et al.  Tyrosine phosphorylation of the fission yeast cdc2+ protein kinase regulates entry into mitosis , 1989, Nature.

[35]  J. Nevins,et al.  Cell cycle regulation of the E2F transcription factor involves an interaction with cyclin A , 1991, Cell.

[36]  A. Vojtek,et al.  Ras-Raf interaction: two-hybrid analysis. , 1995, Methods in enzymology.

[37]  T. Hunter,et al.  The differential localization of human cyclins A and B is due to a cytoplasmic retention signal in cyclin B. , 1994, The EMBO journal.

[38]  Tony Hunter,et al.  Isolation of a human cyclin cDNA: Evidence for cyclin mRNA and protein regulation in the cell cycle and for interaction with p34cdc2 , 1989, Cell.

[39]  D. Beach,et al.  The fission yeast cdc2/cdc13/suc1 protein kinase: Regulation of catalytic activity and nuclear localization , 1989, Cell.

[40]  J. Moore,et al.  Control of cyclin B1 localization through regulated binding of the nuclear export factor CRM1. , 1998, Genes & development.

[41]  K. Gould,et al.  Phosphorylation at Thr167 is required for Schizosaccharomyces pombe p34cdc2 function. , 1991, The EMBO journal.

[42]  R. Kraft,et al.  Two different subunits of importin cooperate to recognize nuclear localization signals and bind them to the nuclear envelope , 1995, Current Biology.

[43]  Jeremy Minshull,et al.  Cyclin is a component of maturation-promoting factor from Xenopus , 1990, Cell.

[44]  J. Labbé,et al.  The MO15 gene encodes the catalytic subunit of a protein kinase that activates cdc2 and other cyclin‐dependent kinases (CDKs) through phosphorylation of Thr161 and its homologues. , 1993, The EMBO journal.

[45]  T. Hunt,et al.  The role of cyclin synthesis, modification and destruction in the control of cell division , 1989, Journal of Cell Science.

[46]  J. Pines,et al.  Temporal and spatial control of cyclin B1 destruction in metaphase , 1999, Nature Cell Biology.

[47]  T. Hunter,et al.  Membrane localization of the kinase which phosphorylates p34cdc2 on threonine 14. , 1994, Molecular biology of the cell.

[48]  E. Nishida,et al.  Nuclear export of cyclin B1 and its possible role in the DNA damage‐induced G2 checkpoint , 1998, The EMBO journal.

[49]  D. Beach,et al.  Activation of cdc2 protein kinase during mitosis in human cells: Cell cycle-dependent phosphorylation and subunit rearrangement , 1988, Cell.

[50]  R. Truant,et al.  Nuclear Import of Cdk/Cyclin Complexes: Identification of Distinct Mechanisms for Import of Cdk2/Cyclin E and Cdc2/Cyclin B1 , 1999, The Journal of cell biology.

[51]  Marc W. Kirschner,et al.  Cyclin activation of p34 cdc2 , 1990, Cell.

[52]  J. Labbé,et al.  MPF from starfish oocytes at first meiotic metaphase is a heterodimer containing one molecule of cdc2 and one molecule of cyclin B. , 1989, The EMBO journal.

[53]  P. Russell,et al.  p80cdc25 mitotic inducer is the tyrosine phosphatase that activates p34cdc2 kinase in fission yeast. , 1991, The EMBO journal.

[54]  M. Kirschner,et al.  Role of phosphorylation in p34cdc2 activation: identification of an activating kinase. , 1992, Molecular biology of the cell.

[55]  E. Hartmann,et al.  Distinct functions for the two importin subunits in nuclear protein import , 1995, Nature.

[56]  D. Donoghue,et al.  Speedy: a novel cell cycle regulator of the G2/M transition , 1999, The EMBO journal.

[57]  A. N. Meyer,et al.  Requirement for phosphorylation of cyclin B1 for Xenopus oocyte maturation. , 1995, Molecular biology of the cell.