Identification and molecular evolution of new dynein-like protein sequences in rat brain

1995 ◽  
Vol 108 (5) ◽  
pp. 1883-1893 ◽  
Author(s):  
Y. Tanaka ◽  
Z. Zhang ◽  
N. Hirokawa
Keyword(s):  
Rat Brain ◽  
Heavy Chain ◽  
Phylogenetic Trees ◽  
Cytoplasmic Dynein ◽  
Amino Acid Sequences ◽  
Adult Brain ◽  
Chain Gene ◽  
Evolutionary Analysis ◽  
Degenerate Primers ◽  
Dynein Heavy Chain

RT-PCR cloning was performed to find unknown members of the dynein superfamily expressed in rat brain. Six kinds of degenerate primers designed for the dynein catalytic domain consensuses were used for extensive PCR amplifications. We have sequenced 550 plasmid clones which turned out to include 13 kinds of new dynein-like sequences (DLP1-8, 9A/B, 10–12) and cytoplasmic dynein heavy chain. In these clones, alternative splicing was detected for a 105 nt-domain containing the CFDEFNRI consensus just downstream of the most N-terminal P-loop (DLP9A and 9B). By using these obtained sequences, initial hybridization studies were performed. Genomic Southern blotting showed each sequence corresponds to a single copy of the gene, while northern blotting of adult brain presented more than one band for some subtypes. We further accomplished molecular evolutionary analysis to recognize their phylogenetic origins for the axonemal and non-axonemal (cytoplasmic) functions. Different methods (UPGMA, NJ and MP) presented well coincident phylogenetic trees from 44 partial amino acid sequences of dynein heavy chain from various eukaryotes. The trunk for all the cytoplasmic dynein heavy chain homologues diverged directly from the root of the phylogenetic tree, suggesting that the first dynein gene duplication defined two distinct functions as respective subfamilies. Of particular interest, we found a duplication event of the cytoplasmic dynein heavy chain gene giving rise to another subtype, DLP4, located between the divergence of yeast and that of Dictyostelium. Such evolutionary topology builds up an inceptive hypothesis that there are at least two non-axonemal dynein heavy chains in mammals.

scholarly journals The primary structure of rat brain (cytoplasmic) dynein heavy chain, a cytoplasmic motor enzyme.

1993 ◽  
Vol 90 (17) ◽  
pp. 7928-7932 ◽  
Author(s):  
Z. Zhang ◽  
Y. Tanaka ◽  
S. Nonaka ◽  
H. Aizawa ◽  
H. Kawasaki ◽  
...  
Keyword(s):  
Rat Brain ◽  
Heavy Chain ◽  
Primary Structure ◽  
Cytoplasmic Dynein ◽  
Dynein Heavy Chain

614 The primary structure of rat brain (cytoplasmic) dynein heavy chain, a cytoplasmic motor enzyme

1993 ◽  
Vol 18 ◽  
pp. S75
Author(s):  
Zhizeng Zhang ◽  
Yosuke Tanaka ◽  
Shigenori Nonaka ◽  
Hiroyuki Aizawa ◽  
Takao Nakata ◽  
...  
Keyword(s):  
Rat Brain ◽  
Heavy Chain ◽  
Primary Structure ◽  
Cytoplasmic Dynein ◽  
Dynein Heavy Chain

scholarly journals A family of dynein genes in Drosophila melanogaster.

1994 ◽  
Vol 5 (1) ◽  
pp. 45-55 ◽  
Author(s):  
K Rasmusson ◽  
M Serr ◽  
J Gepner ◽  
I Gibbons ◽  
T S Hays
Keyword(s):  
Amino Acid ◽  
Heavy Chain ◽  
Amino Acid Level ◽  
Single Copy ◽  
Cytoplasmic Dynein ◽  
Amino Acid Identity ◽  
Chain Gene ◽  
Phosphate Binding ◽  
Atp Binding Site ◽  
Dynein Heavy Chain

We report the identification and initial characterization of seven Drosophila dynein heavy chain genes. Each gene is single copy and maps to a unique genomic location. Sequence analysis of partial clones reveals that each encodes a highly conserved portion of the putative dynein hydrolytic ATP-binding site in dyneins that includes a consensus phosphate-binding (P-loop) motif. One of the clones is derived from a Drosophila cytoplasmic dynein heavy chain gene, Dhc64C, that shows extensive amino acid identity to cytoplasmic dynein isoforms from other organisms. Two other Drosophila dynein clones are 85 and 90% identical at the amino acid level to the corresponding region of the beta heavy chain of sea urchin axonemal dynein. Probes for all seven of the dynein-related sequences hybridize to transcripts that are of the appropriate size, approximately 14 kilobases, to encode the characteristic high molecular weight dynein heavy chain polypeptides. The Dhc64C transcript is readily detected in RNA from ovaries, embryos, and testes. Transcripts from five of the six remaining genes are also detected in much lesser amounts in tissues other than testes. All but one of the dynein transcripts are expressed at comparable levels in testes suggesting their participation in flagellar axoneme assembly and motility.

Cytoplasmic Dynein Function Is Essential in Drosophila melanogaster

Genetics ◽  
1996 ◽  
Vol 142 (3) ◽  
pp. 865-878 ◽  
Author(s):  
Janice Gepner ◽  
Min-gang Li ◽  
Susan Ludmann ◽  
Cynthia Kortas ◽  
Kristin Boylan ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Intracellular Transport ◽  
Mutational Analysis ◽  
Cytoplasmic Dynein ◽  
Complementation Group ◽  
Transport Processes ◽  
Female Sterility ◽  
Chain Gene ◽  
Heavy Chain Gene ◽  
Dynein Heavy Chain

Abstract The microtubule motor cytoplasmic dynein has been implicated in a variety of intracellular transport processes. We previously identified and characterized the Drosophila gene Dhc64C, which encodes a cytoplasmic dynein heavy chain. To investigate the function of the cytoplasmic dynein motor, we initiated a mutational analysis of the Dhc64C dynein gene. A small deletion that removes the chromosomal region containing the heavy chain gene was used to isolate EMS-induced lethal mutations that define at least eight essential genes in the region. Germline transformation with a Dhc64C transgene rescued 16 mutant alleles in the single complementation group that identifies the dynein heavy chain gene. All 16 alleles were hemizygous lethal, which demonstrates that the cytoplasmic dynein heavy chain gene Dhc64C is essential for Drosophila development. Furthermore, our failure to recover somatic clones of cells homozygous for a Dhc64C mutation indicates that cytoplasmic dynein function is required for cell viability in several Drosophila tissues. The intragenic complementation of dynein alleles reveals multiple mutant phenotypes including male and/or female sterility, bristle defects, and defects in eye development.

scholarly journals Dynein from Dictyostelium: primary structure comparisons between a cytoplasmic motor enzyme and flagellar dynein.

1992 ◽  
Vol 119 (6) ◽  
pp. 1597-1604 ◽  
Author(s):  
M P Koonce ◽  
P M Grissom ◽  
J R McIntosh
Keyword(s):  
Sea Urchin ◽  
Heavy Chain ◽  
Sequence Similarity ◽  
Consensus Sequence ◽  
Cytoplasmic Dynein ◽  
Chain Gene ◽  
Cellular Slime Mold ◽  
Reading Frame ◽  
Dynein Heavy Chain ◽  
Binding Domains

We report here the cloning and sequencing of a cytoplasmic dynein heavy chain gene from the cellular slime mold Dictyostelium discoideum. Using a combination of approaches, we have isolated 14,318 bp of DNA sequence which contains an open-reading frame of 4,725 amino acids. The deduced molecular weight of the polypeptide predicted by this reading frame is 538,482 D. Overall, the polypeptide sequence is 51% similar and 28% identical to the recently published sequences of the beta-dynein heavy chain from sea urchin flagella (Gibbons, I. R., B. H. Gibbons, G. Mocz, and D. J. Asai. 1991. Nature (Lond.). 352: 640-643; Ogawa, K. 1991. Nature (Lond.). 352:643-645). It contains four GXXXXGKT/S motifs that form part of a consensus sequence for ATP-binding domains; these motifs are clustered near the middle of the polypeptide. The distribution of the regions sharing sequence similarity between the Dictyostelium and sea urchin heavy chain polypeptides suggests that the amino termini of dyneins may contain domains that specify axonemal or cytoplasmic functions.

scholarly journals Cytoplasmic Dynein Is Required for the Nuclear Attachment and Migration of Centrosomes during Mitosis in Drosophila

1999 ◽  
Vol 146 (3) ◽  
pp. 597-608 ◽  
Author(s):  
John T. Robinson ◽  
Edward J. Wojcik ◽  
Mark A. Sanders ◽  
Maura McGrail ◽  
Thomas S. Hays
Keyword(s):  
Heavy Chain ◽  
Spatial Organization ◽  
Critical Role ◽  
Genetic System ◽  
Cytoplasmic Dynein ◽  
Chain Gene ◽  
Wild Type ◽  
Dynein Heavy Chain ◽  
And Migration

Cytoplasmic dynein is a multisubunit minus-end–directed microtubule motor that serves multiple cellular functions. Genetic studies in Drosophila and mouse have demonstrated that dynein function is essential in metazoan organisms. However, whether the essential function of dynein reflects a mitotic requirement, and what specific mitotic tasks require dynein remains controversial. Drosophila is an excellent genetic system in which to analyze dynein function in mitosis, providing excellent cytology in embryonic and somatic cells. We have used previously characterized recessive lethal mutations in the dynein heavy chain gene, Dhc64C, to reveal the contributions of the dynein motor to mitotic centrosome behavior in the syncytial embryo. Embryos lacking wild-type cytoplasmic dynein heavy chain were analyzed by in vivo analysis of rhodamine-labeled microtubules, as well as by immu-nofluorescence in situ methods. Comparisons between wild-type and Dhc64C mutant embryos reveal that dynein function is required for the attachment and migration of centrosomes along the nuclear envelope during interphase/prophase, and to maintain the attachment of centrosomes to mitotic spindle poles. The disruption of these centrosome attachments in mutant embryos reveals a critical role for dynein function and centrosome positioning in the spatial organization of the syncytial cytoplasm of the developing embryo.

scholarly journals Cytoplasmic Dynein Is Required for Distinct Aspects of Mtoc Positioning, Including Centrosome Separation, in the One Cell Stage Caenorhabditis elegans Embryo

1999 ◽  
Vol 147 (1) ◽  
pp. 135-150 ◽  
Author(s):  
Pierre Gönczy ◽  
Silke Pichler ◽  
Matthew Kirkham ◽  
Anthony A. Hyman
Keyword(s):  
Caenorhabditis Elegans ◽  
Heavy Chain ◽  
Cytoplasmic Dynein ◽  
Chain Gene ◽  
Differential Interference Contrast Microscopy ◽  
Microtubule Organizing Center ◽  
Cell Stage ◽  
Dynein Heavy Chain ◽  
Centrosome Separation ◽  
The One

We have investigated the role of cytoplasmic dynein in microtubule organizing center (MTOC) positioning using RNA-mediated interference (RNAi) in Caenorhabditis elegans to deplete the product of the dynein heavy chain gene dhc-1. Analysis with time-lapse differential interference contrast microscopy and indirect immunofluorescence revealed that pronuclear migration and centrosome separation failed in one cell stage dhc-1 (RNAi) embryos. These phenotypes were also observed when the dynactin components p50/dynamitin or p150Glued were depleted with RNAi. Moreover, in 15% of dhc-1 (RNAi) embryos, centrosomes failed to remain in proximity of the male pronucleus. When dynein heavy chain function was diminished only partially with RNAi, centrosome separation took place, but orientation of the mitotic spindle was defective. Therefore, cytoplasmic dynein is required for multiple aspects of MTOC positioning in the one cell stage C. elegans embryo. In conjunction with our observation of cytoplasmic dynein distribution at the periphery of nuclei, these results lead us to propose a mechanism in which cytoplasmic dynein anchored on the nucleus drives centrosome separation.

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