Cyclin Cln3 is retained at the ER and released by the J chaperone Ydj1 in late G1 to trigger cell cycle entry.

G1 cyclin Cln3 plays a key role in linking cell growth and proliferation in budding yeast. It is generally assumed that Cln3, which is present throughout G1, accumulates passively in the nucleus until a threshold is reached to trigger cell cycle entry. We show here that Cln3 is retained bound to the ER in early G1 cells. ER retention requires binding of Cln3 to the cyclin-dependent kinase Cdc28, a fraction of which also associates to the ER. Cln3 contains a chaperone-regulatory Ji domain that counteracts Ydj1, a J chaperone essential for ER release and nuclear accumulation of Cln3 in late G1. Finally, Ydj1 is limiting for release of Cln3 and timely entry into the cell cycle. As protein synthesis and ribosome assembly rates compromise chaperone availability, we hypothesize that Ydj1 transmits growth capacity information to the cell cycle for setting efficient size/ploidy ratios.

[1]  S. Landry,et al.  Role of the J-domain in the cooperation of Hsp40 with Hsp70. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

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

[3]  J. Brodsky,et al.  Nucleotide Exchange Factor for the Yeast Hsp70 Molecular Chaperone Ssa1p , 2002, Molecular and Cellular Biology.

[4]  C. Wittenberg,et al.  CLN3, not positive feedback, determines the timing of CLN2 transcription in cycling cells. , 1995, Genes & development.

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

[6]  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.

[7]  E. Garí,et al.  The Cln3 cyclin is down‐regulated by translational repression and degradation during the G1 arrest caused by nitrogen deprivation in budding yeast , 1997, The EMBO journal.

[8]  B. Futcher,et al.  Relationship between the function and the location of G1 cyclins in S. cerevisiae. , 2001, Journal of cell science.

[9]  Mike Tyers,et al.  How Cells Coordinate Growth and Division , 2004, Current Biology.

[10]  F. Cross,et al.  Identification of novel and conserved functional and structural elements of the G1 cyclin Cln3 important for interactions with the CDK Cdc28 in Saccharomyces cerevisiae , 2005, Yeast.

[11]  M. Polymenis,et al.  Bem1p, a scaffold signaling protein, mediates cyclin-dependent control of vacuolar homeostasis in Saccharomyces cerevisiae. , 2005, Genes & development.

[12]  M Aldea,et al.  A Set of Vectors with a Tetracycline‐Regulatable Promoter System for Modulated Gene Expression in Saccharomyces cerevisiae , 1997, Yeast.

[13]  Laura L. Newcomb,et al.  AZF1 Is a Glucose-Dependent Positive Regulator of CLN3 Transcription in Saccharomyces cerevisiae , 2002, Molecular and Cellular Biology.

[14]  K. Nasmyth At the heart of the budding yeast cell cycle. , 1996, Trends in genetics : TIG.

[15]  I. Stansfield,et al.  An MBoC Favorite: TOR controls translation initiation and early G1 progression in yeast , 2012, Molecular biology of the cell.

[16]  Mike Tyers,et al.  CDK Activity Antagonizes Whi5, an Inhibitor of G1/S Transcription in Yeast , 2004, Cell.

[17]  B. Futcher,et al.  p34Cdc28-mediated control of Cln3 cyclin degradation , 1995, Molecular and cellular biology.

[18]  B. Carter,et al.  Genes which control cell proliferation in the yeast Saccharomyces cerevisiae , 1980, Nature.

[19]  F. Cross,et al.  DAF1, a mutant gene affecting size control, pheromone arrest, and cell cycle kinetics of Saccharomyces cerevisiae , 1988, Molecular and cellular biology.

[20]  Curt Wittenberg,et al.  Cln3 Activates G1-Specific Transcription via Phosphorylation of the SBF Bound Repressor Whi5 , 2004, Cell.

[21]  B. Futcher,et al.  Isolation and characterization of WHI3, a size-control gene of Saccharomyces cerevisiae. , 2001, Genetics.

[22]  B. Edgar,et al.  Why size matters: altering cell size. , 2002, Current opinion in genetics & development.

[23]  F. Cross,et al.  The yeast Cln3 protein is an unstable activator of Cdc28 , 1993, Molecular and cellular biology.

[24]  B. Futcher Transcriptional regulatory networks and the yeast cell cycle. , 2002, Current opinion in cell biology.

[25]  A. Emili,et al.  Hsc70 Regulates Accumulation of Cyclin D1 and Cyclin D1-Dependent Protein Kinase , 2003, Molecular and Cellular Biology.

[26]  Carol A. Gross,et al.  Structural Features Required for the Interaction of the Hsp70 Molecular Chaperone DnaK with Its Cochaperone DnaJ* , 1999, The Journal of Biological Chemistry.

[27]  S. Reed,et al.  Subcellular localization of a protein kinase required for cell cycle initiation in Saccharomyces cerevisiae: evidence for an association between the CDC28 gene product and the insoluble cytoplasmic matrix , 1987, The Journal of cell biology.

[28]  T. Lithgow,et al.  The J‐protein family: modulating protein assembly, disassembly and translocation , 2004, EMBO reports.

[29]  C. Georgopoulos,et al.  The Conserved G/F Motif of the DnaJ Chaperone Is Necessary for the Activation of the Substrate Binding Properties of the DnaK Chaperone (*) , 1995, The Journal of Biological Chemistry.

[30]  Frederick R. Cross,et al.  Mechanisms Controlling Subcellular Localization of the G1 Cyclins Cln2p and Cln3p in Budding Yeast , 2001, Molecular and Cellular Biology.

[31]  Michael Ruogu Zhang,et al.  Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. , 1998, Molecular biology of the cell.

[32]  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.

[33]  J. Brodsky,et al.  Specific molecular chaperone interactions and an ATP-dependent conformational change are required during posttranslational protein translocation into the yeast ER. , 1998, Molecular biology of the cell.

[34]  L. Breeden,et al.  A novel Mcm1-dependent element in the SWI4, CLN3, CDC6, and CDC47 promoters activates M/G1-specific transcription. , 1997, Genes & development.

[35]  S. Jentsch,et al.  Mobilization of Processed, Membrane-Tethered SPT23 Transcription Factor by CDC48UFD1/NPL4, a Ubiquitin-Selective Chaperone , 2001, Cell.

[36]  E. Craig,et al.  Functional interaction of cytosolic hsp70 and a DnaJ-related protein, Ydj1p, in protein translocation in vivo , 1996, Molecular and cellular biology.

[37]  P. Nurse,et al.  Growth in cell length in the fission yeast Schizosaccharomyces pombe. , 1985, Journal of cell science.

[38]  Trevor Lithgow,et al.  Protein hijacking: key proteins held captive against their will. , 2004, Cancer cell.

[39]  J. Mitchison The growth of single cells. II. Saccharomyces cerevisiae. , 1958, Experimental cell research.

[40]  L. Hartwell,et al.  Genetic control of the cell division cycle in yeast. , 1974, Science.

[41]  M. Polymenis,et al.  Coupling of cell division to cell growth by translational control of the G1 cyclin CLN3 in yeast. , 1997, Genes & development.

[42]  D. Cyr,et al.  YDJ1p facilitates polypeptide translocation across different intracellular membranes by a conserved mechanism , 1992, Cell.

[43]  B. Andrews,et al.  Regulation of Cell Cycle Transcription Factor Swi4 through Auto-Inhibition of DNA Binding , 1999, Molecular and Cellular Biology.

[44]  M. Kasten,et al.  3 p and Rpd 3 p transcriptional regulators . A large protein complex containing the yeast , 1996 .

[45]  P. Novick,et al.  Dynamics and inheritance of the endoplasmic reticulum , 2004, Journal of Cell Science.

[46]  Jian Zhang,et al.  Genomic Scale Mutant Hunt Identifies Cell Size Homeostasis Genes in S. cerevisiae , 2002, Current Biology.

[47]  G. Tokiwa,et al.  The WHI1+ gene of Saccharomyces cerevisiae tethers cell division to cell size and is a cyclin homolog. , 1988, The EMBO journal.

[48]  M Aldea,et al.  Whi3 binds the mRNA of the G1 cyclin CLN3 to modulate cell fate in budding yeast. , 2001, Genes & development.

[49]  N. Pfanner,et al.  The Presequence Translocase-associated Protein Import Motor of Mitochondria , 2004, Journal of Biological Chemistry.

[50]  Soojin Lee,et al.  Exchangeable chaperone modules contribute to specification of type I and type II Hsp40 cellular function. , 2003, Molecular biology of the cell.

[51]  Frank Holstege,et al.  Predicting gene function through systematic analysis and quality assessment of high-throughput data , 2005, Bioinform..

[52]  C. Woldringh,et al.  Volume growth of daughter and parent cells during the cell cycle of Saccharomyces cerevisiae a/alpha as determined by image cytometry , 1993, Journal of bacteriology.

[53]  Bruce Futcher,et al.  Growth Rate and Cell Size Modulate the Synthesis of, and Requirement for, G1-Phase Cyclins at Start , 2004, Molecular and Cellular Biology.

[54]  M. Tyers,et al.  A dynamic transcriptional network communicates growth potential to ribosome synthesis and critical cell size. , 2004, Genes & development.

[55]  E. Garí,et al.  Recruitment of Cdc28 by Whi3 restricts nuclear accumulation of the G1 cyclin–Cdk complex to late G1 , 2004, The EMBO journal.

[56]  A. Goldberg,et al.  The molecular chaperone Ydj1 is required for the p34CDC28-dependent phosphorylation of the cyclin Cln3 that signals its degradation , 1996, Molecular and cellular biology.

[57]  C. Kaiser,et al.  Physiological Regulation of Membrane Protein Sorting Late in the Secretory Pathway of Saccharomyces cerevisiae , 1997, The Journal of cell biology.

[58]  Mike Tyers,et al.  Systematic Identification of Pathways That Couple Cell Growth and Division in Yeast , 2002, Science.

[59]  B. Novák,et al.  The size control of fission yeast revisited. , 1996, Journal of cell science.

[60]  G C Johnston,et al.  Coordination of growth with cell division in the yeast Saccharomyces cerevisiae. , 1977, Experimental cell research.