The Saccharomyces cerevisiae Start-specific transcription factor Swi4 interacts through the ankyrin repeats with the mitotic Clb2/Cdc28 kinase and through its conserved carboxy terminus with Swi6

At a point in late G1 termed Start, yeast cells enter S phase, duplicate their spindle pole bodies, and form buds. These events require activation of Cdc28 kinase by G1 cyclins. Swi4 associates with Swi6 to form the SCB-binding factor complex which activates G1 cyclin genes CLN1 and CLN2 in late G1. In G2 and M phases, the transcriptional activity of SCB-binding factor is repressed by the mitotic Clb2/Cdc28 kinase. Mbp1, a transcription factor related to Swi4, forms the MCB-binding factor complex with Swi6, which activates DNA synthesis genes and S-phase cyclin genes CLB5 and CLB6 in late G1. Clb2/Cdc28 kinase is not required for the repression of MCB-binding factor transcriptional activity in G2 and M phase. We show here that the Swi4 carboxy terminus is sufficient for interaction with Swi6 in vitro. A carboxy-terminal domain of Swi6 is required and sufficient for interaction with Swi4. The carboxy terminus of Mbp1 is sufficient for interaction with Swi6, and the carboxy terminus of Swi6 is required for interaction with Mbp1. By coimmunoprecipitation, we show that Swi4 but not Mbp1 interacts with Clb2/Cdc28 kinase in vivo during the G2 and M phases of the cell cycle. We demonstrate that the ankyrin repeats of Swi4 mediate the interaction with Clb2/Cdc28 kinase. The ankyrin repeats constitute a domain by which a cell cycle-specific transcription factor can interact with cyclin-dependent kinase complexes, thus enabling it to link its transcriptional activity to cell cycle progression.

[1]  L. Moore,et al.  Mutational analysis of a DNA sequence involved in linking gene expression to the cell cycle. , 1992, Biochemistry and cell biology = Biochimie et biologie cellulaire.

[2]  J. Decaprio,et al.  The Schizosaccharomyces pombe MBF complex requires heterodimerization for entry into S phase , 1995, Molecular and cellular biology.

[3]  Kim Nasmyth,et al.  The role of SWI4 and SWI6 in the activity of G1 cyclins in yeast , 1991, Cell.

[4]  B. Futcher,et al.  Comparison of the Saccharomyces cerevisiae G1 cyclins: Cln3 may be an upstream activator of Cln1, Cln2 and other cyclins. , 1993, The EMBO journal.

[5]  S. Reed,et al.  The role of p34 kinases in the G1 to S-phase transition. , 1992, Annual review of cell biology.

[6]  B. Dynlacht,et al.  Differential regulation of E2F transactivation by cyclin/cdk2 complexes. , 1994, Genes & development.

[7]  L. Dirick,et al.  Roles and regulation of Cln‐Cdc28 kinases at the start of the cell cycle of Saccharomyces cerevisiae. , 1995, The EMBO journal.

[8]  D. Smith,et al.  Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. , 1988, Gene.

[9]  K Nasmyth,et al.  CLB5 and CLB6, a new pair of B cyclins involved in DNA replication in Saccharomyces cerevisiae. , 1993, Genes & development.

[10]  S. Forsburg,et al.  The fission yeast cdc18 + gene product couples S phase to START and mitosis , 1993, Cell.

[11]  Uttam Surana,et al.  The role of CDC28 and cyclins during mitosis in the budding yeast S. cerevisiae , 1991, Cell.

[12]  C. Peng,et al.  Cyclin A/CDK2 binds directly to E2F-1 and inhibits the DNA-binding activity of E2F-1/DP-1 by phosphorylation , 1994, Molecular and cellular biology.

[13]  Andrew J. Bannister,et al.  The activation domain of transcription factor PU.1 binds the retinoblastoma (RB) protein and the transcription factor TFIID in vitro: RB shows sequence similarity to TFIID and TFIIB. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[14]  I. Herskowitz,et al.  Transcriptional activation of CLN1, CLN2, and a putative new G1 cyclin (HCS26) by SWI4, a positive regulator of G1-specific transcription. , 1991, Cell.

[15]  S. Reed,et al.  Cyclin-B homologs in Saccharomyces cerevisiae function in S phase and in G2. , 1992, Genes & development.

[16]  L. Johnston,et al.  Control of DNA synthesis genes in fission yeast by the cell-cycle gene cdclO + , 1992, Nature.

[17]  Michael Primig,et al.  Anatomy of a transcription factor important for the Start of the cell cycle in Saccharomyces cerevisiae , 1992, Nature.

[18]  Wilhelm Krek,et al.  Negative regulation of the growth-promoting transcription factor E2F-1 by a stably bound cyclin A-dependent protein kinase , 1994, Cell.

[19]  Daniel J. Lew,et al.  A cyclin B homolog in S. cerevisiae: Chronic activation of the Cdc28 protein kinase by cyclin prevents exit from mitosis , 1991, Cell.

[20]  K Nasmyth,et al.  Control of the yeast cell cycle by the Cdc28 protein kinase. , 1993, Current opinion in cell biology.

[21]  K Nasmyth,et al.  Switching transcription on and off during the yeast cell cycle: Cln/Cdc28 kinases activate bound transcription factor SBF (Swi4/Swi6) at start, whereas Clb/Cdc28 kinases displace it from the promoter in G2. , 1996, Genes & development.

[22]  Kim Nasmyth,et al.  Cell cycle control of the yeast HO gene: Cis- and Trans-acting regulators , 1987, Cell.

[23]  I. Herskowitz,et al.  Cell cycle control by a complex of the cyclin HCS26 (PCL1) and the kinase PHO85. , 1994, Science.

[24]  F. Cross.  Cell cycle arrest caused by CLN gene deficiency in Saccharomyces cerevisiae resembles START-I arrest and is independent of the mating-pheromone signalling pathway , 1990, Molecular and cellular biology.

[25]  Gregory J. Hannon,et al.  pl5INK4B is a potentia| effector of TGF-β-induced cell cycle arrest , 1994, Nature.

[26]  Kim Nasmyth,et al.  A central role for SWI6 in modulating cell cycle Start-specific transcription in yeast , 1992, Nature.

[27]  D. Beach,et al.  cdt1 is an essential target of the Cdc10/Sct1 transcription factor: requirement for DNA replication and inhibition of mitosis. , 1994, The EMBO journal.

[28]  R. D. Gietz,et al.  New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. , 1988, Gene.

[29]  C. O'keefe,et al.  Growth suppression by p18, a p16INK4/MTS1- and p14INK4B/MTS2-related CDK6 inhibitor, correlates with wild-type pRb function. , 1994, Genes & development.

[30]  Kim Nasmyth,et al.  A repetitive DNA sequence that confers cell-cycle START (CDC28)-dependent transcription of the HO gene in yeast , 1985, Cell.

[31]  A. Pardee.  G1 events and regulation of cell proliferation. , 1989, Science.

[32]  K. Tanaka,et al.  A new cdc gene required for S phase entry of Schizosaccharomyces pombe encodes a protein similar to the cdc 10+ and SWI4 gene products. , 1992, The EMBO journal.

[33]  Mike Tyers,et al.  Mechanisms that help the yeast cell cycle clock tick: G2 cyclins transcriptionally activate G2 cyclins and repress G1 cyclins , 1993, Cell.

[34]  L. Breeden,et al.  Similarity between cell-cycle genes of budding yeast and fission yeast and the Notch gene of Drosophila , 1987, Nature.

[35]  E. O’Shea,et al.  Phosphate-regulated inactivation of the kinase PHO80-PHO85 by the CDK inhibitor PHO81. , 1994, Science.

[36]  N. Ogawa,et al.  Functional domains of Pho81p, an inhibitor of Pho85p protein kinase, in the transduction pathway of Pi signals in Saccharomyces cerevisiae , 1995, Molecular and cellular biology.

[37]  K. Tanaka,et al.  res2+, a new member of the cdc10+/SWI4 family, controls the ‘start’ of mitotic and meiotic cycles in fission yeast. , 1994, EMBO Journal.

[38]  D. Beach,et al.  Sct1 functions in partnership with Cdc10 in a transcription complex that activates cell cycle START and inhibits differentiation , 1993, Cell.

[39]  M. Tyers,et al.  The PCL2 (ORFD)-PHO85 cyclin-dependent kinase complex: a cell cycle regulator in yeast. , 1994, Science.

[40]  K. Nasmyth,et al.  A role for the transcription factors Mbp1 and Swi4 in progression from G1 to S phase. , 1993, Science.

[41]  P. Bork.  Hundreds of ankyrin‐like repeats in functionally diverse proteins: Mobile modules that cross phyla horizontally? , 1993, Proteins.

[42]  L. Johnston,et al.  Coordination of expression of DNA synthesis genes in budding yeast by a cell-cycle regulated trans factor , 1991, Nature.

[43]  N. L. Thangue,et al.  DP and E2F proteins: components of a heterodimeric transcription factor implicated in cell cycle control , 1994 .

[44]  I. Herskowitz,et al.  Identification of a DNA binding factor involved in cell-cycle control of the yeast HO gene , 1989, Cell.

[45]  K. Nasmyth,et al.  Changes in a SWI4,6-DNA-binding complex occur at the time of HO gene activation in yeast. , 1991, Genes & development.

[46]  L. Breeden,et al.  Analysis of the SWI4/SWI6 protein complex, which directs G1/S-specific transcription in Saccharomyces cerevisiae , 1993, Molecular and cellular biology.

[47]  L. Moore,et al.  Interaction of the yeast Swi4 and Swi6 cell cycle regulatory proteins in vitro. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[48]  The inhibitor of DNA replication encoded by the Drosophila gene plutonium is a small, ankyrin repeat protein. , 1994, The EMBO journal.

[49]  B. Futcher,et al.  The Cln3‐Cdc28 kinase complex of S. cerevisiae is regulated by proteolysis and phosphorylation. , 1992, The EMBO journal.

[50]  G. Hannon,et al.  A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4 , 1993, Nature.

[51]  F. Cross,et al.  CLB5: a novel B cyclin from budding yeast with a role in S phase. , 1992, Genes & development.

[52]  Curt Wittenberg,et al.  An essential G1 function for cyclin-like proteins in yeast , 1989, Cell.