Characterization of the KLP68D kinesin-like protein in Drosophila: possible roles in axonal transport.

This paper describes the molecular and biochemical properties of KLP68D, a new kinesin-like motor protein in Drosophila melanogaster. Sequence analysis of a full-length cDNA encoding KLP68D demonstrates that this protein has a domain that shares significant sequence identity with the entire 340-amin acid kinesin heavy chain motor domain. Sequences extending beyond the motor domain predict a region of alpha-helical coiled-coil followed by a globular "tail" region; there is significant sequence similarity between the alpha-helical coiled-coil region of the KLP68D protein and similar regions of the KIF3 protein of mouse and the KRP85 protein of sea urchin. This finding suggests that all three proteins may be members of the same family, and that they all perform related functions. KLP68D protein produced in Escherichia coli is, like kinesin itself, a plus-end directed microtubule motor. In situ hybridization analysis of KLP68D RNA in Drosophila embryos indicates that the KLP68D gene is expressed primarily in the central nervous system and in a subset of the peripheral nervous system during embryogenesis. Thus, KLP68D may be used for anterograde axonal transport and could conceivably move cargoes in fly neurons different than those moved by kinesin heavy chain or other plus-end directed motors.

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Kinesin family in murine central nervous system.

The Journal of Cell Biology ◽

10.1083/jcb.119.5.1287 ◽

1992 ◽

Vol 119 (5) ◽

pp. 1287-1296 ◽

Cited By ~ 197

Author(s):

H Aizawa ◽

Y Sekine ◽

R Takemura ◽

Z Zhang ◽

M Nangaku ◽

...

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Amino Acid ◽

Amino Acid Sequence ◽

Heavy Chain ◽

Motor Domain ◽

Globular Domain ◽

Complete Amino Acid Sequence ◽

Kinesin Heavy Chain ◽

Kinesin Family

In neuronal axons, various kinds of membranous components are transported along microtubules bidirectionally. However, only two kinds of mechanochemical motor proteins, kinesin and brain dynein, had been identified as transporters of membranous organelles in mammalian neurons. Recently, a series of genes that encode proteins closely related to kinesin heavy chain were identified in several organisms including Schizosaccharomyces pombe, Aspergillus niddulans, Saccharomyces cerevisiae, Caenorhabditus elegans, and Drosophila. Most of these members of the kinesin family are implicated in mechanisms of mitosis or meiosis. To address the mechanism of intracellular organelle transport at a molecular level, we have cloned and characterized five different members (KIF1-5), that encode the microtubule-associated motor domain homologous to kinesin heavy chain, in murine brain tissue. Homology analysis of amino acid sequence indicated that KIF1 and KIF5 are murine counterparts of unc104 and kinesin heavy chain, respectively, while KIF2, KIF3, and KIF4 are as yet unidentified new species. Complete amino acid sequence of KIF3 revealed that KIF3 consists of NH2-terminal motor domain, central alpha-helical rod domain, and COOH-terminal globular domain. Complete amino acid sequence of KIF2 revealed that KIF2 consists of NH2-terminal globular domain, central motor domain, and COOH-terminal alpha-helical rod domain. This is the first identification of the kinesin-related protein which has its motor domain at the central part in its primary structure. Northern blot analysis revealed that KIF1, KIF3, and KIF5 are expressed almost exclusively in murine brain, whereas KIF2 and KIF4 are expressed in brain as well as in other tissues. All these members of the kinesin family are expressed in the same type of neurons, and thus each one of them may transport its specific organelle in the murine central nervous system.

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Partially Functional Outer-Arm Dynein in a Novel Chlamydomonas Mutant Expressing a Truncated γ Heavy Chain

Eukaryotic Cell ◽

10.1128/ec.00102-08 ◽

2008 ◽

Vol 7 (7) ◽

pp. 1136-1145 ◽

Cited By ~ 36

Author(s):

Zhongmei Liu ◽

Hiroko Takazaki ◽

Yuki Nakazawa ◽

Miho Sakato ◽

Toshiki Yagi ◽

...

Keyword(s):

Heavy Chain ◽

Motor Domain ◽

Basal Region ◽

Terminal Sequence ◽

Tail Region ◽

Heavy Chains ◽

Thin Section Electron Microscopy ◽

Section Electron ◽

And Function ◽

Necessary And Sufficient

ABSTRACT The outer dynein arm of Chlamydomonas flagella contains three heavy chains (α, β, and γ), each of which exhibits motor activity. How they assemble and cooperate is of considerable interest. Here we report the isolation of a novel mutant, oda2-t, whose γ heavy chain is truncated at about 30% of the sequence. While the previously isolated γ chain mutant oda2 lacks the entire outer arm, oda2-t retains outer arms that contain α and β heavy chains, suggesting that the N-terminal sequence (corresponding to the tail region) is necessary and sufficient for stable outer-arm assembly. Thin-section electron microscopy and image analysis localize the γ heavy chain to a basal region of the outer-arm image in the axonemal cross section. The motility of oda2-t is lower than that of the wild type and oda11 (lacking the α heavy chain) but higher than that of oda2 and oda4-s7 (lacking the motor domain of the β heavy chain). Thus, the outer-arm dynein lacking the γ heavy-chain motor domain is partially functional. The availability of mutants lacking individual heavy chains should greatly facilitate studies on the structure and function of the outer-arm dynein.

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Structural and biochemical properties of kinesin heavy chain associated with rat brain mitochondria

Cell Motility and the Cytoskeleton ◽

10.1002/cm.970280108 ◽

1994 ◽

Vol 28 (1) ◽

pp. 79-93 ◽

Cited By ~ 25

Author(s):

Abdeljelil Jellali ◽

Marie-H�l�ne Metz-Boutigue ◽

Irina Surgucheva ◽

Veronika Jancsik ◽

Christian Schwartz ◽

...

Keyword(s):

Rat Brain ◽

Heavy Chain ◽

Brain Mitochondria ◽

Biochemical Properties ◽

Rat Brain Mitochondria ◽

Kinesin Heavy Chain

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Expression of Neuronal Kinesin Heavy Chain Is Developmentally Regulated in the Central Nervous System of the Rat

Journal of Neurochemistry ◽

10.1046/j.1471-4159.1997.69051840.x ◽

2002 ◽

Vol 69 (5) ◽

pp. 1840-1849 ◽

Cited By ~ 8

Author(s):

G. Vignali ◽

C. Lizier ◽

M. T. Sprocati ◽

C. Sirtori ◽

G. Battaglia ◽

...

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Heavy Chain ◽

Developmentally Regulated ◽

Kinesin Heavy Chain ◽

The Central Nervous System

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Cloning and expression of a human kinesin heavy chain gene: interaction of the COOH-terminal domain with cytoplasmic microtubules in transfected CV-1 cells

The Journal of Cell Biology ◽

10.1083/jcb.117.6.1263 ◽

1992 ◽

Vol 117 (6) ◽

pp. 1263-1275 ◽

Cited By ~ 121

Author(s):

F Navone ◽

J Niclas ◽

N Hom-Booher ◽

L Sparks ◽

HD Bernstein ◽

...

Keyword(s):

Heavy Chain ◽

Translational Regulation ◽

Coiled Coil ◽

Gene Interaction ◽

Cloning And Expression ◽

Upstream Open Reading Frame ◽

Chain Gene ◽

Terminal Domain ◽

Kinesin Heavy Chain ◽

Cytoplasmic Microtubules

To understand the interactions between the microtubule-based motor protein kinesin and intracellular components, we have expressed the kinesin heavy chain and its different domains in CV-1 monkey kidney epithelial cells and examined their distributions by immunofluorescence microscopy. For this study, we cloned and sequenced cDNAs encoding a kinesin heavy chain from a human placental library. The human kinesin heavy chain exhibits a high level of sequence identity to the previously cloned invertebrate kinesin heavy chains; homologies between the COOH-terminal domain of human and invertebrate kinesins and the nonmotor domain of the Aspergillus kinesin-like protein bimC were also found. The gene encoding the human kinesin heavy chain also contains a small upstream open reading frame in a G-C rich 5' untranslated region, features that are associated with translational regulation in certain mRNAs. After transient expression in CV-1 cells, the kinesin heavy chain showed both a diffuse distribution and a filamentous staining pattern that coaligned with microtubules but not vimentin intermediate filaments. Altering the number and distribution of microtubules with taxol or nocodazole produced corresponding changes in the localization of the expressed kinesin heavy chain. The expressed NH2-terminal motor and the COOH-terminal tail domains, but not the alpha-helical coiled coil rod domain, also colocalized with microtubules. The finding that both the kinesin motor and tail domains can interact with cytoplasmic microtubules raises the possibility that kinesin could crossbridge and induce sliding between microtubules under certain circumstances.

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Multiple actin-based motor genes in Dictyostelium.

Cell Regulation ◽

10.1091/mbc.1.1.55 ◽

1989 ◽

Vol 1 (1) ◽

pp. 55-63 ◽

Cited By ~ 66

Author(s):

M A Titus ◽

H M Warrick ◽

J A Spudich

Keyword(s):

Molecular Motors ◽

Myosin Head ◽

Sequence Similarity ◽

Coiled Coil ◽

Large Family ◽

Helical Structure ◽

Amino Acid Sequences ◽

Tail Region ◽

Head Region ◽

Amino Terminal

Dictyostelium cells, devoid of conventional myosin, display a variety of motile activities, consistent with the presence of other molecular motors. The Dictyostelium genome was probed at low stringency with a gene fragment containing the conserved conventional myosin head domain sequences to identify other actin-based motors that may play a role in the observed motility of these mutant cells. One gene (abmA) has been characterized and encodes a polypeptide of approximately 135 kDa with a head region homologous to other myosin head sequences and a tail region that is not predicted to form either an alpha-helical structure of coiled-coil interactions. Comparisons of the amino acid sequences of the tail regions of abmA, Dictyostelium myosin I, and Acanthamoeba myosins IB and IL reveal an area of sequence similarity in the amino terminal half of the tail that may be a membrane-binding domain. The abmA gene, however, does not contain an unusual Gly, Pro, Ala stretch typical of many of the previously described myosin Is. Two additional genes (abmB and abmC) were identified using this approach and also found to contain sequences that encode proteins with typical conserved myosin head sequences. The abm genes may be part of a large family of actin-based motors that play various roles in diverse aspects of cellular motility.

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Fast axonal transport of kinesin in the rat visual system: functionality of kinesin heavy chain isoforms.

Molecular Biology of the Cell ◽

10.1091/mbc.6.1.21 ◽

1995 ◽

Vol 6 (1) ◽

pp. 21-40 ◽

Cited By ~ 79

Author(s):

R G Elluru ◽

G S Bloom ◽

S T Brady

Keyword(s):

Axonal Transport ◽

Heavy Chain ◽

Slow Axonal Transport ◽

Fast Axonal Transport ◽

Anterograde Transport ◽

Kinesin Heavy Chain ◽

Order Of Magnitude ◽

Fast Transport ◽

Regulation Of Transport ◽

Cytoplasmic Structures

The mechanochemical ATPase kinesin is thought to move membrane-bounded organelles along microtubules in fast axonal transport. However, fast transport includes several classes of organelles moving at rates that differ by an order of magnitude. Further, the fact that cytoplasmic forms of kinesin exist suggests that kinesins might move cytoplasmic structures such as the cytoskeleton. To define cellular roles for kinesin, the axonal transport of kinesin was characterized. Retinal proteins were pulse-labeled, and movement of radiolabeled kinesin through optic nerve and tract into the terminals was monitored by immunoprecipitation. Heavy and light chains of kinesin appeared in nerve and tract at times consistent with fast transport. Little or no kinesin moved with slow axonal transport indicating that effectively all axonal kinesin is associated with membranous organelles. Both kinesin heavy chain molecular weight variants of 130,000 and 124,000 M(r) (KHC-A and KHC-B) moved in fast anterograde transport, but KHC-A moved at 5-6 times the rate of KHC-B. KHC-A cotransported with the synaptic vesicle marker synaptophysin, while a portion of KHC-B cotransported with the mitochondrial marker hexokinase. These results suggest that KHC-A is enriched on small tubulovesicular structures like synaptic vesicles and that at least one form of KHC-B is predominantly on mitochondria. Biochemical specialization may target kinesins to appropriate organelles and facilitate differential regulation of transport.

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