Structure of the acrosomal bundle

In the unactivated Limulus sperm, a 60-µm-long bundle of actin filaments crosslinked by the protein scruin is bent and twisted into a coil around the base of the nucleus. At fertilization, the bundle uncoils and fully extends in five seconds to support a finger of membrane known as the acrosomal process. This biological spring is powered by stored elastic energy and does not require the action of motor proteins or actin polymerization. In a 9.5-Å electron cryomicroscopic structure of the extended bundle, we show that twist, tilt and rotation of actin–scruin subunits deviate widely from a ‘standard’ F-actin filament. This variability in structural organization allows filaments to pack into a highly ordered and rigid bundle in the extended state and suggests a mechanism for storing and releasing energy between coiled and extended states without disassembly.

[1]  L. G. Tilney,et al.  Actin filaments in the acrosomal reaction of Limulus sperm. Motion generated by alterations in the packing of the filaments , 1975, The Journal of cell biology.

[2]  D. DeRosier,et al.  A change in the twist of the actin-containing filaments occurs during the extension of the acrosomal process in Limulus sperm. , 1980, Journal of molecular biology.

[3]  D. DeRosier,et al.  F-actin is a helix with a random variable twist , 1982, Nature.

[4]  G. Lucchini,et al.  A short nucleotide sequence required for regulation of HIS4 by the general control system of yeast , 1983, Cell.

[5]  M. Ptashne,et al.  The carboxy-terminal 30 amino acids of GAL4 are recognized by GAL80 , 1987, Cell.

[6]  D J DeRosier,et al.  Three-dimensional reconstruction of an actin bundle , 1988, The Journal of cell biology.

[7]  B. Errede,et al.  STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast, is part of protein-DNA complexes. , 1989, Genes & development.

[8]  W. Kabsch,et al.  Atomic structure of the actin: DNase I complex , 1990, Nature.

[9]  W. Kabsch,et al.  Atomic model of the actin filament , 1990, Nature.

[10]  T. D. Schneider,et al.  Sequence logos: a new way to display consensus sequences. , 1990, Nucleic acids research.

[11]  B. Cochran,et al.  The PRE and PQ box are functionally distinct yeast pheromone response elements , 1990, Molecular and cellular biology.

[12]  G. Fink,et al.  Functional redundancy in the yeast cell cycle: FUS3 and KSS1 have both overlapping and unique functions. , 1991, Cold Spring Harbor symposia on quantitative biology.

[13]  G. Fink,et al.  FUS3 represses CLN1 and CLN2 and in concert with KSS1 promotes signal transduction. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[14]  S. Fields,et al.  Pheromone-dependent phosphorylation of the yeast STE12 protein correlates with transcriptional activation. , 1991, Genes & development.

[15]  M. Primig,et al.  The DNA binding and oligomerization domain of MCM1 is sufficient for its interaction with other regulatory proteins. , 1991, The EMBO journal.

[16]  M. Brandriss,et al.  Proline-independent binding of PUT3 transcriptional activator protein detected by footprinting in vivo , 1991, Molecular and cellular biology.

[17]  K. Nasmyth,et al.  Signal transduction in Saccharomyces cerevisiae requires tyrosine and threonine phosphorylation of FUS3 and KSS1. , 1992, Genes & development.

[18]  W. Chiu,et al.  Imaging frozen, hydrated acrosomal bundle from Limulus sperm at 7 A resolution with a 400 kV electron cryomicroscope. , 1993, Journal of molecular biology.

[19]  G. Fink,et al.  Elements of the yeast pheromone response pathway required for filamentous growth of diploids. , 1993, Science.

[20]  G. Fink,et al.  A role for autophosphorylation revealed by activated alleles of FUS3, the yeast MAP kinase homolog. , 1994, Molecular biology of the cell.

[21]  R. KNÜPPEL,et al.  TRANSFAC Retrieval Program: A Network Model Database of Eukaryotic Transcription Regulating Sequences and Proteins , 1994, J. Comput. Biol..

[22]  W. Timberlake,et al.  The Aspergillus nidulans abaA gene encodes a transcriptional activator that acts as a genetic switch to control development , 1994, Molecular and cellular biology.

[23]  G. Fink,et al.  Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth. , 1994, Genes & development.

[24]  G. Fink,et al.  Regulated degradation of the transcription factor Gcn4. , 1994, The EMBO journal.

[25]  J Jakana,et al.  Three-dimensional structure of a single filament in the Limulus acrosomal bundle: scruin binds to homologous helix-loop-beta motifs in actin , 1994, The Journal of cell biology.

[26]  A. Futcher,et al.  Use of polymerase chain reaction epitope tagging for protein tagging in Saccharomyces cerevisiae , 1995, Yeast.

[27]  P. Schimmel,et al.  Detection of leucine-independent DNA site occupancy of the yeast Leu3p transcriptional activator in vivo , 1995, Molecular and cellular biology.

[28]  P. Matsudaira,et al.  Sequence and domain organization of scruin, an actin-cross-linking protein in the acrosomal process of Limulus sperm , 1995, The Journal of cell biology.

[29]  J. Dickinson,et al.  'Fusel' alcohols induce hyphal-like extensions and pseudohyphal formation in yeasts. , 1996, Microbiology.

[30]  L. Bardwell,et al.  Two novel targets of the MAP kinase Kss1 are negative regulators of invasive growth in the yeast Saccharomyces cerevisiae. , 1996, Genes & development.

[31]  Paul Matsudaira,et al.  Characterization of the Actin Cross-linking Properties of the Scruin-Calmodulin Complex from the Acrosomal Process of Limulus Sperm (*) , 1996, The Journal of Biological Chemistry.

[32]  G. Fink,et al.  Combinatorial Control Required for the Specificity of Yeast MAPK Signaling , 1997, Science.

[33]  B. Errede,et al.  Cooperative binding interactions required for function of the Ty1 sterile responsive element , 1997, Molecular and cellular biology.

[34]  S. Fields,et al.  Transcriptional activation upon pheromone stimulation mediated by a small domain of Saccharomyces cerevisiae Ste12p , 1997, Molecular and cellular biology.

[35]  Wah Chiu,et al.  Cofilin Changes the Twist of F-Actin: Implications for Actin Filament Dynamics and Cellular Function , 1997, The Journal of cell biology.

[36]  M. Tyers,et al.  Regulation of the mating pheromone and invasive growth responses in yeast by two MAP kinase substrates , 1997, Current Biology.

[37]  L. Bardwell,et al.  Inhibitory and activating functions for MAPK Kss1 in the S. cerevisiae filamentous- growth signalling pathway , 1997, Nature.

[38]  B. Wasylyk,et al.  Ets transcription factors: nuclear effectors of the Ras-MAP-kinase signaling pathway. , 1998, Trends in biochemical sciences.

[39]  G. Church,et al.  Finding DNA regulatory motifs within unaligned noncoding sequences clustered by whole-genome mRNA quantitation , 1998, Nature Biotechnology.

[40]  G. Fink,et al.  The riddle of MAP kinase signaling specificity. , 1998, Trends in genetics : TIG.

[41]  L. Bardwell,et al.  Differential regulation of transcription: repression by unactivated mitogen-activated protein kinase Kss1 requires the Dig1 and Dig2 proteins. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Michael Q. Zhang,et al.  SCPD: a promoter database of the yeast Saccharomyces cerevisiae , 1999, Bioinform..

[43]  James I. Garrels,et al.  The Yeast Proteome Database (YPD): a model for the organization and presentation of genome-wide functional data , 1999, Nucleic Acids Res..

[44]  E. Goldsmith,et al.  Relative dependence of different outputs of the Saccharomyces cerevisiae pheromone response pathway on the MAP kinase Fus3p. , 1999, Genetics.

[45]  Michael N. Hall,et al.  The TOR signalling pathway controls nuclear localization of nutrient-regulated transcription factors , 1999, Nature.

[46]  J Jakana,et al.  The three-dimensional structure of the Limulus acrosomal process: a dynamic actin bundle. , 1999, Journal of molecular biology.

[47]  R. Wisdom,et al.  AP-1: one switch for many signals. , 1999, Experimental cell research.

[48]  S. Fields,et al.  Constitutive activation of the Saccharomyces cerevisiae transcriptional regulator Ste12p by mutations at the amino‐terminus , 2000, Yeast.

[49]  T. Hughes,et al.  Signaling and circuitry of multiple MAPK pathways revealed by a matrix of global gene expression profiles. , 2000, Science.

[50]  John J. Wyrick,et al.  Genome-wide location and function of DNA binding proteins. , 2000, Science.

[51]  M. Simon,et al.  Receptor Tyrosine Kinases Specific Outcomes from General Signals , 2000, Cell.

[52]  L. Mahadevan,et al.  Motility powered by supramolecular springs and ratchets. , 2000, Science.

[53]  Ivan Sadowski,et al.  Two Regulators of Ste12p Inhibit Pheromone-Responsive Transcription by Separate Mechanisms , 2000, Molecular and Cellular Biology.

[54]  E. Elion,et al.  Pheromone response, mating and cell biology. , 2000, Current opinion in microbiology.

[55]  W. Sabbagh,et al.  Specificity of MAP kinase signaling in yeast differentiation involves transient versus sustained MAPK activation. , 2001, Molecular cell.

[56]  Nicola J. Rinaldi,et al.  Serial Regulation of Transcriptional Regulators in the Yeast Cell Cycle , 2001, Cell.

[57]  S. Kron,et al.  Cell cycle control of yeast filamentous growth. , 2001, Current opinion in microbiology.

[58]  MAP kinases bite back. , 2001, Developmental cell.

[59]  M. Baker,et al.  Bridging the information gap: computational tools for intermediate resolution structure interpretation. , 2001, Journal of molecular biology.

[60]  David Botstein,et al.  Promoter-specific binding of Rap1 revealed by genome-wide maps of protein–DNA association , 2001, Nature Genetics.

[61]  E. Egelman,et al.  Probing the structure of F-actin: cross-links constrain atomic models and modify actin dynamics. , 2001, Journal of molecular biology.

[62]  M. Rose,et al.  Yeast mating: Getting close to membrane merger , 2001, Current Biology.

[63]  Willy Wriggers,et al.  Actin Depolymerizing Factor Stabilizes an Existing State of F-Actin and Can Change the Tilt of F-Actin Subunits , 2001, The Journal of cell biology.

[64]  Edward H. Egelman,et al.  Actin allostery again? , 2001, Nature Structural Biology.

[65]  M. Cobb,et al.  MAP kinases. , 2001, Chemical reviews.

[66]  L. Otterbein,et al.  The Crystal Structure of Uncomplexed Actin in the ADP State , 2001, Science.

[67]  M. Karin,et al.  Mammalian MAP kinase signalling cascades , 2001, Nature.

[68]  M. Peter,et al.  MAP Kinase Cascades: Scaffolding Signal Specificity , 2002, Current Biology.

[69]  Edward H. Egelman,et al.  The utrophin actin-binding domain binds F-actin in two different modes , 2002, The Journal of cell biology.

[70]  P. Lazarovici,et al.  Signaling Pathways for PC12 Cell Differentiation: Making the Right Connections , 2002, Science.

[71]  Nicola J. Rinaldi,et al.  Transcriptional Regulatory Networks in Saccharomyces cerevisiae , 2002, Science.

[72]  M. Snyder,et al.  Bud-site selection and cell polarity in budding yeast. , 2002, Current opinion in microbiology.

[73]  M. Tyers,et al.  MAPK signaling specificity: it takes two to tango. , 2002, Trends in cell biology.

[74]  Edward H. Egelman,et al.  How does ATP hydrolysis control actin's associations? , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[75]  Edward H. Egelman,et al.  The bacterial protein SipA polymerizes G-actin and mimics muscle nebulin , 2002, Nature Structural Biology.

[76]  M. Duterque-Coquillaud,et al.  When Ets transcription factors meet their partners. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[77]  A. Michelson Deciphering genetic regulatory codes: A challenge for functional genomics , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[78]  Ting Wang,et al.  Combining phylogenetic data with co-regulated genes to identify regulatory motifs , 2003, Bioinform..

[79]  L. Fulton,et al.  Finding Functional Features in Saccharomyces Genomes by Phylogenetic Footprinting , 2003, Science.

[80]  Roberto Dominguez,et al.  Crystal Structure of Monomeric Actin in the ATP State , 2003, Journal of Biological Chemistry.

[81]  L. Mahadevan,et al.  Stored elastic energy powers the 60-m extension of the Limulus polyphemus sperm actin bundle , 2003 .

[82]  Mathieu Blanchette,et al.  FootPrinter: a program designed for phylogenetic footprinting , 2003, Nucleic Acids Res..

[83]  Roberto Dominguez,et al.  CRYSTAL STRUCTURE OF MONOMERIC ACTIN IN THE ATP STATE , 2003 .

[84]  B. Birren,et al.  Sequencing and comparison of yeast species to identify genes and regulatory elements , 2003, Nature.

[85]  Belinda Bullard,et al.  The location of ubiquitin in Lethocerus arthrin. , 2003, Journal of molecular biology.

[86]  Rasmus R. Schröder,et al.  Electron cryo-microscopy shows how strong binding of myosin to actin releases nucleotide , 2003, Nature.

[87]  Roberto Dominguez,et al.  Solution properties of TMR-actin: when biochemical and crystal data agree. , 2003, Biophysical journal.

[88]  Michael F Schmid,et al.  Cross-correlation and merging of crystallographic reflections derived from cryoelectron micrographs of 3D crystals: application to the Limulus acrosomal bundle. , 2003, Journal of structural biology.

[89]  D. Weitz,et al.  Elastic Behavior of Cross-Linked and Bundled Actin Networks , 2004, Science.

[90]  P. Matsudaira,et al.  Bending stiffness of a crystalline actin bundle. , 2004, Journal of molecular biology.

[91]  C G dos Remedios,et al.  Fluorescence depolarization of actin filaments in reconstructed myofibers: the effect of S1 or pPDM-S1 on movements of distinct areas of actin. , 2004, Biophysical journal.

[92]  Michael A. Beer,et al.  Whole-genome discovery of transcription factor binding sites by network-level conservation. , 2003, Genome research.