glsA, a Volvox gene required for asymmetric division and germ cell specification, encodes a chaperone-like protein.

The gls genes of Volvox are required for the asymmetric divisions that set apart cells of the germ and somatic lineages during embryogenesis. Here we used transposon tagging to clone glsA, and then showed that it is expressed maximally in asymmetrically dividing embryos, and that it encodes a 748-amino acid protein with two potential protein-binding domains. Site-directed mutagenesis of one of these, the J domain (by which Hsp40-class chaperones bind to and activate specific Hsp70 partners) abolishes the capacity of glsA to rescue mutants. Based on this and other considerations, including the fact that the GlsA protein is associated with the mitotic spindle, we discuss how it might function, in conjunction with an Hsp70-type partner, to shift the division plane in asymmetrically dividing cells.

[1]  P. Silver,et al.  A yeast DnaJ homologue, Scj1p, can function in the endoplasmic reticulum with BiP/Kar2p via a conserved domain that specifies interactions with Hsp70s , 1995, The Journal of cell biology.

[2]  K. Arndt,et al.  The yeast SIS1 protein, a DnaJ homolog, is required for the initiation of translation , 1993, Cell.

[3]  H. Saibil,et al.  T-complex polypeptide-1 is a subunit of a heteromeric particle in the eukaryotic cytosol , 1992, Nature.

[4]  M. Culbertson,et al.  The yeast homolog to mouse Tcp-1 affects microtubule-mediated processes , 1991, Molecular and cellular biology.

[5]  P. Benfey,et al.  The SCARECROW Gene Regulates an Asymmetric Cell Division That Is Essential for Generating the Radial Organization of the Arabidopsis Root , 1996, Cell.

[6]  John G. White,et al.  The dynactin complex is required for cleavage plane specification in early Caenorhabditis elegans embryos , 1998, Current Biology.

[7]  C. Georgopoulos,et al.  Isolation and characterization of dnaJ null mutants of Escherichia coli , 1990, Journal of bacteriology.

[8]  J. Cooper,et al.  Transient localized accumulation of actin in Caenorhabditis elegans blastomeres with oriented asymmetric divisions. , 1994, Development.

[9]  M. Kirschner,et al.  Systematic identification of mitotic phosphoproteins , 1997, Current Biology.

[10]  J. Ahringer,et al.  G Proteins Are Required for Spatial Orientation of Early Cell Cleavages in C. elegans Embryos , 1996, Cell.

[11]  M. Douglas,et al.  A Conserved HPD Sequence of the J-domain Is Necessary for YDJ1 Stimulation of Hsp70 ATPase Activity at a Site Distinct from Substrate Binding (*) , 1996, The Journal of Biological Chemistry.

[12]  C. Doe Spindle Orientation and Asymmetric Localization in Drosophila: Both Inscuteable? , 1996, Cell.

[13]  D. Kirk,et al.  The program for cellular differentiation in Volvox carteri as revealed by molecular analysis of development in a gonidialess/somatic regenerator mutant. , 1991, Development.

[14]  R. Webster,et al.  Localization, synthesis, and activity of an antigenic site on influenza virus hemagglutinin. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[15]  C. Doe,et al.  Extrinsic cues, intrinsic cues and microfilaments regulate asymmetric protein localization in Drosophila neuroblasts , 1997, Current Biology.

[16]  R. C. Starr Structure, reproduction and differentiation in Volvox carteri f. nagariensis Iyengar, Strains HK 9 & 10 , 1969 .

[17]  D. Kirk,et al.  Genetic and cytological control of the asymmetric divisions that pattern the Volvox embryo. , 1991, Development (Cambridge, England). Supplement.

[18]  A. Ransick,et al.  The relationship between cell size and cell fate in Volvox carteri , 1993, The Journal of cell biology.

[19]  E. Craig,et al.  2 Cytosolic hsp70s of Saccharomyces cerevisiae : Roles in Protein Synthesis, Protein Translocation, Proteolysis, and Regulation , 1994 .

[20]  S. Dutcher,et al.  Loss of spatial control of the mitotic spindle apparatus in a Chlamydomonas reinhardtii mutant strain lacking basal bodies. , 1995, Genetics.

[21]  D. Kirk,et al.  In search of molecular origins of cellular differentiation in Volvox and its relatives. , 1992, International review of cytology.

[22]  W. Shoji,et al.  MIDA1, a Protein Associated with Id, Regulates Cell Growth (*) , 1995, The Journal of Biological Chemistry.

[23]  T. Hays,et al.  The microtubule motor cytoplasmic dynein is required for spindle orientation during germline cell divisions and oocyte differentiation in Drosophila. , 1997, Development.

[24]  D. Cyr,et al.  Eukaryotic homologues of Escherichia coli dnaJ: a diverse protein family that functions with hsp70 stress proteins. , 1993, Molecular biology of the cell.

[25]  G. Kochert,et al.  SPERM BUNDLE‐FEMALE SOMATIC CELL INTERACTION IN THE FERTILIZATION PROCESS OF VOLVOX CARTERI F. WEISMANNIA(CHLOROPHYTA) 1 , 1979 .

[26]  M. Wigler,et al.  Purification of a RAS-responsive adenylyl cyclase complex from Saccharomyces cerevisiae by use of an epitope addition method , 1988, Molecular and cellular biology.

[27]  S. Dutcher,et al.  Pharmacological and genetic evidence for a role of rootlet and phycoplast microtubules in the positioning and assembly of cleavage furrows in Chlamydomonas reinhardtii. , 1998, Cell motility and the cytoskeleton.

[28]  D. Kirk,et al.  Jordan, an active Volvox transposable element similar to higher plant transposons. , 1993, The Plant cell.

[29]  Y. Jan,et al.  Role of inscuteable in orienting asymmetric cell divisions in Drosophila , 1996, Nature.

[30]  C. Lloyd The Cytoskeletal basis of plant growth and form , 1991 .

[31]  K. Kemphues,et al.  A non-muscle myosin required for embryonic polarity in Caenorhabditis elegans , 1996, Nature.

[32]  P. Silver,et al.  Eukaryotic DnaJ homologs and the specificity of Hsp70 activity , 1993, Cell.

[33]  W. Welch,et al.  Molecular Chaperones and the Centrosome , 1996, The Journal of Biological Chemistry.

[34]  Ira Herskowitz,et al.  Mechanisms of asymmetric cell division: Two Bs or not two Bs, that is the question , 1992, Cell.

[35]  R. Quatrano,et al.  Fucus Embryogenesis: A Model to Study the Establishment of Polarity. , 1993, The Plant cell.

[36]  S. Strome,et al.  Cleavage Plane Specification in C. elegans: How to Divide the Spoils , 1996, Cell.

[37]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[38]  S. Lindquist,et al.  Role of the protein chaperone YDJ1 in establishing Hsp90-mediated signal transduction pathways. , 1995, Science.

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

[40]  R. Starr,et al.  Control of differentiation in Volvox. , 1970, The ... Symposium. Society for Developmental Biology. Symposium.

[41]  M. Bate,et al.  Development of the indirect flight muscles of Drosophila. , 1991, Development.

[42]  S. Strome Determination of cleavage planes , 1993, Cell.

[43]  D. Kirk,et al.  Translational regulation of protein synthesis, in response to light, at a critical stage of volvox development , 1985, Cell.

[44]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[45]  W. Müller,et al.  Nuclear transformation of Volvox carteri. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[46]  T. Langer,et al.  DnaJ-like proteins: molecular chaperones and specific regulators of Hsp70. , 1994, Trends in biochemical sciences.

[47]  L. Gerace,et al.  Identification of novel M phase phosphoproteins by expression cloning. , 1996, Molecular biology of the cell.

[48]  K. Johnson,et al.  Identification of a molecular chaperone in the eukaryotic flagellum and its localization to the site of microtubule assembly. , 1995, Journal of cell science.

[49]  R. Schekman,et al.  A Sec63p-BiP complex from yeast is required for protein translocation in a reconstituted proteoliposome , 1993, The Journal of cell biology.

[50]  W. Kelley,et al.  The J-domain family and the recruitment of chaperone power. , 1998, Trends in biochemical sciences.

[51]  J. Raff,et al.  Centrosomes, and not nuclei, initiate pole cell formation in Drosophila embryos , 1989, Cell.