The Saccharomyces cerevisiae SDA1 gene is required for actin cytoskeleton organization and cell cycle progression.

The organization of the actin cytoskeleton is essential for several cellular processes. Here we report the characterization of a Saccharomyces cerevisiae novel gene, SDA1, encoding a highly conserved protein, which is essential for cell viability and is localized in the nucleus. Depletion or inactivation of Sda1 cause cell cycle arrest in G(1) by blocking both budding and DNA replication, without loss of viability. Furthermore, sda1-1 temperature-sensitive mutant cells arrest at the non-permissive temperature mostly without detectable structures of polymerized actin, although a normal actin protein level is maintained, indicating that Sda1 is required for proper organization of the actin cytoskeleton. To our knowledge, this is the first mutation shown to cause such a phenotype. Recovery of Sda1 activity restores proper assembly of actin structures, as well as budding and DNA replication. Furthermore we show that direct actin perturbation, either in sda1-1 or in cdc28-13 cells released from G(1) block, prevents recovery of budding and DNA replication. We also show that the block in G(1) caused by loss of Sda1 function is independent of Swe1. Altogether our results suggest that disruption of F-actin structure can block cell cycle progression in G(1) and that Sda1 is involved in the control of the actin cytoskeleton.

[1]  M. Kikyo,et al.  Bni1p Regulates Microtubule-Dependent Nuclear Migration through the Actin Cytoskeleton in Saccharomyces cerevisiae , 1999, Molecular and Cellular Biology.

[2]  Chandra L. Theesfeld,et al.  The Morphogenesis Checkpoint in Saccharomyces cerevisiae: Cell Cycle Control of Swe1p Degradation by Hsl1p and Hsl7p , 1999, Molecular and Cellular Biology.

[3]  D. Amberg,et al.  Three-dimensional imaging of the yeast actin cytoskeleton through the budding cell cycle. , 1998, Molecular biology of the cell.

[4]  Daniel J. Lew,et al.  A Morphogenesis Checkpoint Monitors the Actin Cytoskeleton in Yeast , 1998, The Journal of cell biology.

[5]  Daniel J. Lew,et al.  Involvement of an Actomyosin Contractile Ring in Saccharomyces cerevisiae Cytokinesis , 1998, The Journal of cell biology.

[6]  K. Ayscough,et al.  In vivo functions of actin-binding proteins. , 1998, Current opinion in cell biology.

[7]  M. Snyder,et al.  Cell polarity and morphogenesis in budding yeast. , 1998, Annual review of microbiology.

[8]  M. Carlier,et al.  Control of Actin Dynamics in Cell Motility , 2022 .

[9]  Navin Pokala,et al.  High Rates of Actin Filament Turnover in Budding Yeast and Roles for Actin in Establishment and Maintenance of Cell Polarity Revealed Using the Actin Inhibitor Latrunculin-A , 1997, The Journal of cell biology.

[10]  G. Carignani,et al.  Analysis of a 17·9 kb Region from Saccharomyces cerevisiae Chromosome VII Reveals the Presence of Eight Open Reading Frames, Including BRF1 (TFIIIB70) and GCN5 Genes , 1997, Yeast.

[11]  D. Lew,et al.  Cdc28 tyrosine phosphorylation and the morphogenesis checkpoint in budding yeast. , 1996, Molecular biology of the cell.

[12]  B. Cairns,et al.  TFG/TAF30/ANC1, a component of the yeast SWI/SNF complex that is similar to the leukemogenic proteins ENL and AF-9 , 1996, Molecular and cellular biology.

[13]  D. Drubin,et al.  ACTIN: general principles from studies in yeast. , 1996, Annual review of cell and developmental biology.

[14]  M. Snyder,et al.  Molecular basis of cell integrity and morphogenesis in Saccharomyces cerevisiae. , 1995, Microbiological reviews.

[15]  S. Reed,et al.  Cell cycle control of morphogenesis in budding yeast. , 1995, Current opinion in genetics & development.

[16]  R. Kornberg,et al.  TFIIF-TAF-RNA polymerase II connection. , 1994, Genes & development.

[17]  D. Drubin,et al.  A nuclear protein with sequence similarity to proteins implicated in human acute leukemias is important for cellular morphogenesis and actin cytoskeletal function in Saccharomyces cerevisiae. , 1994, Molecular biology of the cell.

[18]  D. Botstein,et al.  Ultrastructure of the yeast actin cytoskeleton and its association with the plasma membrane , 1994, The Journal of cell biology.

[19]  G. Lucchini,et al.  The B subunit of the DNA polymerase alpha-primase complex in Saccharomyces cerevisiae executes an essential function at the initial stage of DNA replication , 1994, Molecular and cellular biology.

[20]  G. Lucchini,et al.  De novo synthesis of budding yeast DNA polymerase α and POLI transcription at the G1/S boundary are not required for entrance into S phase , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[21]  Gerald R. Fink,et al.  Guide to yeast genetics and molecular biology , 1993 .

[22]  H. Riezman,et al.  Actin and fimbrin are required for the internalization step of endocytosis in yeast. , 1993, The EMBO journal.

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

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

[25]  R. Sikorski,et al.  In vitro mutagenesis and plasmid shuffling: from cloned gene to mutant yeast. , 1991, Methods in enzymology.

[26]  J. Pringle.  Staining of bud scars and other cell wall chitin with calcofluor. , 1991, Methods in enzymology.

[27]  R. Rothstein.  Targeting, disruption, replacement, and allele rescue: integrative DNA transformation in yeast. , 1991, Methods in enzymology.

[28]  H. Riezman,et al.  Yeast endocytosis assays. , 1991, Methods in enzymology.

[29]  J. Pringle,et al.  Staining of actin with fluorochrome-conjugated phalloidin. , 1991, Methods in enzymology.

[30]  R. Laskey,et al.  Nuclear targeting sequences--a consensus? , 1991, Trends in biochemical sciences.

[31]  Fred Winston,et al.  Methods in Yeast Genetics: A Laboratory Course Manual , 1990 .

[32]  S. Reed,et al.  Mitotic role for the Cdc28 protein kinase of Saccharomyces cerevisiae. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Pringle,et al.  CDC42 and CDC43, two additional genes involved in budding and the establishment of cell polarity in the yeast Saccharomyces cerevisiae , 1990, The Journal of cell biology.

[34]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[35]  D Botstein,et al.  Suppressors of yeast actin mutations. , 1989, Genetics.

[36]  B. Haarer,et al.  Fluorescence microscopy methods for yeast. , 1989, Methods in cell biology.

[37]  David Botstein,et al.  Phenotypic Analysis of Temperature-sensitive Yeast Actin Mutants , 2022 .

[38]  R. W. Davis,et al.  A physical, genetic and transcriptional map of the cloned his3 gene region of Saccharomyces cerevisiae. , 1980, Journal of molecular biology.

[39]  D. Botstein,et al.  Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. , 1979, Gene.

[40]  E. Cabib,et al.  Timing and function of chitin synthesis in yeast , 1975, Journal of bacteriology.

[41]  M. Hayashibe,et al.  INITIATION OF BUDDING AND CHITIN-RING , 1973 .