The Neurospora organelle motor: a distant relative of conventional kinesin with unconventional properties.

The "conventional" kinesins comprise a conserved family of molecular motors for organelle transport that have been identified in various animal species. Organelle motors from other phyla have not yet been analyzed at the molecular level. Here we report the identification, biochemical and immunological characterization, and molecular cloning of a cytoplasmic motor in a "lower" eukaryote, the Ascomycete fungus Neurospora crassa. This motor, termed Nkin (for Neurospora kinesin), exhibits several unique structural and functional features, including a high rate of microtubule transport, a lack of copurifying light chains, a second P-loop motif, and an overall sequence organization reminiscent of a kinesin-like protein. However, a greater than average sequence homology in the motor domain and the presence of a highly conserved region in the C-terminus identify Nkin as a distant relative of the family of conventional kinesins. A molecular phylogenetic analysis suggests Nkin to have diverged early in the evolution of this family of motors. The discovery of Nkin may help identify domains important for specific biological functions in conventional kinesins.

[1]  N. Hirokawa,et al.  KIF1B, a novel microtubule plus end-directed monomeric motor protein for transport of mitochondria , 1994, Cell.

[2]  M. McNiven,et al.  Association of kinesin with the Golgi apparatus in rat hepatocytes. , 1994, Journal of cell science.

[3]  H. Goodson,et al.  Molecular phylogeny of the kinesin family of microtubule motor proteins. , 1994, Journal of cell science.

[4]  J. Scholey,et al.  The carboxyl-terminal domain of kinesin heavy chain is important for membrane binding. , 1994, The Journal of biological chemistry.

[5]  Gaston H. Gonnet,et al.  New Algorithms for the Computation of Evolutionary Phylogenetic Trees , 1994 .

[6]  M. Sogin,et al.  Monophyletic origin of animals: a shared ancestry with the fungi. , 1994, Society of General Physiologists series.

[7]  S. Beushausen,et al.  Kinesin light chains: identification and characterization of a family of proteins from the optic lobe of the squid Loligo pealii. , 1993, DNA and cell biology.

[8]  J. Scholey,et al.  Novel heterotrimeric kinesin-related protein purified from sea urchin eggs , 1993, Nature.

[9]  R. Vallee Molecular analysis of the microtubule motor dynein. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[10]  G. Steinberg,et al.  Organelle movements in the wild type and wall-less fz;sg;os-1 mutants of Neurospora crassa are mediated by cytoplasmic microtubules. , 1993, Journal of cell science.

[11]  L. Goldstein,et al.  The Drosophila kinesin light chain. Primary structure and interaction with kinesin heavy chain. , 1993, The Journal of biological chemistry.

[12]  S. Stickel,et al.  Monophyletic origins of the metazoa: an evolutionary link with fungi , 1993, Science.

[13]  U. Plessmann,et al.  Characterization of the tubulin-tyrosine ligase , 1993, The Journal of cell biology.

[14]  P. Meluh,et al.  Suppression of the bimC4 mitotic spindle defect by deletion of klpA, a gene encoding a KAR3-related kinesin-like protein in Aspergillus nidulans , 1993, The Journal of cell biology.

[15]  R. Vallee,et al.  Microtubule and axoneme gliding assays for force production by microtubule motor proteins. , 1993, Methods in cell biology.

[16]  L. Goldstein,et al.  With apologies to scheherazade: tails of 1001 kinesin motors. , 1993, Annual review of genetics.

[17]  J. McIntosh,et al.  Dynein from Dictyostelium: primary structure comparisons between a cytoplasmic motor enzyme and flagellar dynein , 1992, The Journal of cell biology.

[18]  N. Hirokawa,et al.  Kinesin family in murine central nervous system , 1992, The Journal of cell biology.

[19]  M. Sheetz,et al.  Kinectin, a major kinesin-binding protein on ER , 1992, The Journal of cell biology.

[20]  K. Loo,et al.  Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly , 1992, The Journal of cell biology.

[21]  D. Hackney,et al.  Kinesin undergoes a 9 S to 6 S conformational transition. , 1992, The Journal of biological chemistry.

[22]  Rainer Fuchs,et al.  CLUSTAL V: improved software for multiple sequence alignment , 1992, Comput. Appl. Biosci..

[23]  R. Baskin,et al.  Isolation of a sea urchin egg kinesin-related protein using peptide antibodies. , 1992, Journal of cell science.

[24]  T. Mitchison,et al.  Evidence for kinesin-related proteins in the mitotic apparatus using peptide antibodies. , 1992, Journal of Cell Science.

[25]  A. McDowall,et al.  Association of kinesin with characterized membrane-bounded organelles. , 1992, Cell motility and the cytoskeleton.

[26]  G. Bloom,et al.  Molecular genetics of kinesin light chains: generation of isoforms by alternative splicing. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[27]  G. Bloom,et al.  Kinesin associates with anterogradely transported membranous organelles in vivo , 1991, The Journal of cell biology.

[28]  Vladimir Gelfand,et al.  Kinesin is responsible for centrifugal movement of pigment granules in melanophores. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[29]  A. Lupas,et al.  Predicting coiled coils from protein sequences , 1991, Science.

[30]  J. Scholey,et al.  Subcellular localization and sequence of sea urchin kinesin heavy chain: evidence for its association with membranes in the mitotic apparatus and interphase cytoplasm. , 1991 .

[31]  S. Endow,et al.  A multimember kinesin gene family in Drosophila. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[32]  I R Gibbons,et al.  Multiple nucleotide-binding sites in the sequence of dynein beta heavy chain. , 1991, Nature.

[33]  G. Bloom,et al.  Purification of kinesin from bovine brain and assay of microtubule-stimulated ATPase activity. , 1991, Methods in enzymology.

[34]  P. R. Sibbald,et al.  The P-loop--a common motif in ATP- and GTP-binding proteins. , 1990, Trends in biochemical sciences.

[35]  Edward D. Salmon,et al.  The Drosophila claret segregation protein is a minus-end directed motor molecule , 1990, Nature.

[36]  P. Hollenbeck,et al.  Radial extension of macrophage tubular lysosomes supported by kinesin , 1990, Nature.

[37]  W. Neupert,et al.  Matrix processing peptidase of mitochondria. Structure-function relationships. , 1990, The Journal of biological chemistry.

[38]  G. Bloom,et al.  A monoclonal antibody against kinesin inhibits both anterograde and retrograde fast axonal transport in squid axoplasm. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[39]  W. Wickner,et al.  The ATPase activity of secA is regulated by acidic phospholipids, secY, and the leader and mature domains of precursor proteins , 1990, Cell.

[40]  R. Vale,et al.  Identification of a kinesin‐like microtubule‐based motor protein in Dictyostelium discoideum. , 1989, EMBO Journal.

[41]  P. Hollenbeck The distribution, abundance and subcellular localization of kinesin , 1989, The Journal of cell biology.

[42]  G. Bloom,et al.  Monoclonal antibodies to kinesin heavy and light chains stain vesicle- like structures, but not microtubules, in cultured cells , 1989, The Journal of cell biology.

[43]  J. Scholey,et al.  Identification of globular mechanochemical heads of kinesin , 1989, Nature.

[44]  R. A. Laymon,et al.  A three-domain structure of kinesin heavy chain revealed by DNA sequence and microtubule binding analyses , 1989, Cell.

[45]  G. Bloom,et al.  Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration , 1989, Cell.

[46]  D. Murphy,et al.  Isolation of a 45-kDa fragment from the kinesin heavy chain with enhanced ATPase and microtubule-binding activities. , 1989, The Journal of biological chemistry.

[47]  G. Bloom,et al.  Copurification of kinesin polypeptides with microtubule-stimulated Mg-ATPase activity and kinetic analysis of enzymatic properties. , 1989, Cell motility and the cytoskeleton.

[48]  M. Frohman,et al.  Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[49]  J. Scholey,et al.  Inhibition of kinesin-driven microtubule motility by monoclonal antibodies to kinesin heavy chains , 1988, The Journal of cell biology.

[50]  M. Sheetz,et al.  The role of kinesin and other soluble factors in organelle movement along microtubules , 1988, The Journal of cell biology.

[51]  A. Ikai,et al.  Purification and characterization of kinesin from bovine adrenal medulla. , 1988, The Journal of biological chemistry.

[52]  S. Dabora,et al.  The microtubule-dependent formation of a tubulovesicular network with characteristics of the ER from cultured cell extracts , 1988, Cell.

[53]  G. Bloom,et al.  Native structure and physical properties of bovine brain kinesin and identification of the ATP-binding subunit polypeptide. , 1988, Biochemistry.

[54]  D. Lipman,et al.  Improved tools for biological sequence comparison. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[55]  M. Koonce,et al.  An ATPase with properties expected for the organelle motor of the giant amoeba, Reticulomyxa , 1988, Nature.

[56]  L. Goldstein,et al.  Isolation and characterization of the gene encoding the heavy chain of Drosophila kinesin. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[57]  E. Raff,et al.  Drosophila kinesin: characterization of microtubule motility and ATPase. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[58]  E. Vaisberg,et al.  The quaternary structure of bovine brain kinesin. , 1988, The EMBO journal.

[59]  R. Vallee,et al.  MAP 1C is a microtubule-activated ATPase which translocates microtubules in vitro and has dynein-like properties , 1987, The Journal of cell biology.

[60]  J. Scholey,et al.  Correlation between the ATPase and microtubule translocating activities of sea urchin egg kinesin , 1987, Nature.

[61]  Vladimir Gelfand,et al.  Bovine brain kinesin is a microtubule-activated ATPase. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[62]  S. Stanley Earth and life through time , 1986 .

[63]  V. Russo,et al.  Isolating RNA is easy and fun. , 1986 .

[64]  G. Witman Isolation of Chlamydomonas flagella and flagellar axonemes. , 1986, Methods in enzymology.

[65]  J. Olmsted,et al.  Analysis of cytoskeletal structures using blot-purified monospecific antibodies. , 1986, Methods in enzymology.

[66]  J. McIntosh,et al.  Identification of kinesin in sea urchin eggs, and evidence for its localization in the mitotic spindle , 1985, Nature.

[67]  E. Mandelkow,et al.  Tubulin domains probed by limited proteolysis and subunit-specific antibodies. , 1985, Journal of molecular biology.

[68]  Michael P. Sheetz,et al.  Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility , 1985, Cell.

[69]  Scott T. Brady,et al.  A novel brain ATPase with properties expected for the fast axonal transport motor , 1985, Nature.

[70]  R. Smith,et al.  Large-scale isolation of the Neurospora plasma membrane H+-ATPase. , 1984, Analytical biochemistry.

[71]  J. Devereux,et al.  A comprehensive set of sequence analysis programs for the VAX , 1984, Nucleic Acids Res..

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

[73]  M. Schliwa,et al.  Structural interaction of cytoskeletal components , 1981, The Journal of cell biology.

[74]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[75]  W. Neupert,et al.  Preparation of Neurospora crassa mitochondria. , 1979, Methods in enzymology.

[76]  M. O. Dayhoff,et al.  22 A Model of Evolutionary Change in Proteins , 1978 .

[77]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[78]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[79]  Extracts , 1869, The Indian medical gazette.