scholarly journals The t Complex Distorter 2 Candidate Gene, Dnahc8, Encodes at Least Two Testis-Specific Axonemal Dynein Heavy Chains That Differ Extensively at Their Amino and Carboxyl Termini

2002 ◽  
Vol 250 (1) ◽  
pp. 24-43 ◽  
Author(s):  
Sadhana A. Samant ◽  
Olugbemiga Ogunkua ◽  
Ling Hui ◽  
John Fossella ◽  
Stephen H. Pilder
Keyword(s):  
Candidate Gene ◽  
Heavy Chains ◽  
T Complex ◽  
Axonemal Dynein ◽  
Dynein Heavy Chains

scholarly journals WDR92 is required for axonemal dynein heavy chain stability in cytoplasm

2019 ◽  
Vol 30 (15) ◽  
pp. 1834-1845 ◽  
Author(s):  
Ramila S. Patel-King ◽  
Miho Sakato-Antoku ◽  
Maya Yankova ◽  
Stephen M. King
Keyword(s):  
Heavy Chain ◽  
Beat Frequency ◽  
Gel Filtration ◽  
Dynein Heavy Chain ◽  
Heavy Chains ◽  
Assembly Factor ◽  
Axonemal Dynein ◽  
Dynein Arms ◽  
Phylogenetic Signature ◽  
Dynein Heavy Chains

WDR92 associates with a prefoldin-like cochaperone complex and known dynein assembly factors. WDR92 has been very highly conserved and has a phylogenetic signature consistent with it playing a role in motile ciliary assembly or activity. Knockdown of WDR92 expression in planaria resulted in ciliary loss, reduced beat frequency and dyskinetic motion of the remaining ventral cilia. We have now identified a Chlamydomonas wdr92 mutant that encodes a protein missing the last four WD repeats. The wdr92-1 mutant builds only ∼0.7-μm cilia lacking both inner and outer dynein arms, but with intact doublet microtubules and central pair. When cytoplasmic extracts prepared by freeze/thaw from a control strain were fractionated by gel filtration, outer arm dynein components were present in several distinct high molecular weight complexes. In contrast, wdr92-1 extracts almost completely lacked all three outer arm heavy chains, while the IFT dynein heavy chain was present in normal amounts. A wdr92-1 tpg1-2 double mutant builds ∼7-μm immotile flaccid cilia that completely lack dynein arms. These data indicate that WDR92 is a key assembly factor specifically required for the stability of axonemal dynein heavy chains in cytoplasm and suggest that cytoplasmic/IFT dynein heavy chains use a distinct folding pathway.

scholarly journals Identification of seven rat axonemal dynein heavy chain genes: expression during ciliated cell differentiation.

1996 ◽  
Vol 7 (1) ◽  
pp. 71-79 ◽  
Author(s):  
K L Andrews ◽  
P Nettesheim ◽  
D J Asai ◽  
L E Ostrowski
Keyword(s):  
Cell Differentiation ◽  
Heavy Chain ◽  
Ciliated Cell ◽  
Northern Analysis ◽  
Ciliated Cells ◽  
Dynein Heavy Chain ◽  
Heavy Chains ◽  
Cells In Culture ◽  
Axonemal Dynein ◽  
Dynein Heavy Chains

Axonemal dyneins are molecular motors that drive the beating of cilia and flagella. We report here the identification and partial cloning of seven unique axonemal dynein heavy chains from rat tracheal epithelial (RTE) cells. Combinations of axonemal-specific and degenerate primers to conserved regions around the catalytic site of dynein heavy chains were used to obtain cDNA fragments of rat dynein heavy chains. Southern analysis indicates that these are single copy genes, with one possible exception, and Northern analysis of RNA from RTE cells shows a transcript of approximately 15 kb for each gene. Expression of these genes was restricted to tissues containing axonemes (trachea, testis, and brain). A time course analysis during ciliated cell differentiation of RTE cells in culture demonstrated that the expression of axonemal dynein heavy chains correlated with the development of ciliated cells, while cytoplasmic dynein heavy chain expression remained constant. In addition, factors that regulate the development of ciliated cells in culture regulated the expression of axonemal dynein heavy chains in a parallel fashion. These are the first mammalian dynein heavy chain genes shown to be expressed specifically in axonemal tissues. Identification of the mechanisms that regulate the cell-specific expression of these axonemal dynein heavy chains will further our understanding of the process of ciliated cell differentiation.

New Axonemal Dynein Heavy Chains from Tetrahymena thermophila

1999 ◽  
Vol 46 (2) ◽  
pp. 147-154 ◽  
Author(s):  
PAULA S. MOBBERLEY ◽  
JEANELL L. SULLIVAN ◽  
STEVEN P. ANGUS ◽  
XUEJUN KONG ◽  
DAVID G. PENNOCK
Keyword(s):  
Tetrahymena Thermophila ◽  
Heavy Chains ◽  
Axonemal Dynein ◽  
Dynein Heavy Chains

scholarly journals ZMYND10 functions in a chaperone relay during axonemal dynein assembly

2017 ◽  
Author(s):  
Girish R Mali ◽  
Patricia Yeyati ◽  
Seiya Mizuno ◽  
Margaret A Keighren ◽  
Petra zur Lage ◽  
...  
Keyword(s):  
Protein Misfolding ◽  
Specific Protein ◽  
Mouse Genetics ◽  
Rational Drug Design ◽  
Heavy Chains ◽  
Dynein Motor ◽  
Ability To Act ◽  
Axonemal Dynein ◽  
Chaperone Complex ◽  
Dynein Heavy Chains

AbstractMolecular chaperones promote the folding and macromolecular assembly of a diverse set of substrate ‘client’ proteins. How the ubiquitous chaperone machinery directs its activities towards a specific set of substrates and whether this selectivity could be targeted for therapeutic intervention is of intense research. Through the use of mouse genetics, imaging and quantitative proteomics we uncover that ZMYND10 is a novel co-chaperone for the FKBP8-HSP90 chaperone complex during the biosynthesis of axonemal dynein heavy chains required for cilia motility. In the absence of ZMYND10, defects in dynein heavy chains trigger broader dynein motor degradation. We show that FKBP8 inhibition phenocopies dynein motor instability in airway cells, and human disease-causing variants of ZMYND10 disrupt its ability to act as FKBP8-HSP90 co-chaperone. Our study indicates that the motile ciliopathy Primary Ciliary Dyskinesia (PCD) should be considered a cell-type specific protein-misfolding disease and opens the potential for rational drug design that could restore specificity to the ubiquitous chaperone apparatus towards dynein subunits.

scholarly journals Molecular analysis of the gamma heavy chain of Chlamydomonas flagellar outer-arm dynein

1994 ◽  
Vol 107 (3) ◽  
pp. 497-506 ◽  
Author(s):  
C.G. Wilkerson ◽  
S.M. King ◽  
G.B. Witman
Keyword(s):  
Heavy Chain ◽  
Coiled Coil ◽  
Cytoplasmic Dynein ◽  
Complete Sequence ◽  
Dynein Heavy Chain ◽  
Heavy Chains ◽  
Microtubule Binding Domain ◽  
Helical Content ◽  
Axonemal Dynein ◽  
Dynein Heavy Chains

We report here the complete sequence of the gamma dynein heavy chain of the outer arm of the Chlamydomonas flagellum, and partial sequences for six other dynein heavy chains. The gamma dynein heavy chain sequence contains four P-loop motifs, one of which is the likely hydrolytic site based on its position relative to a previously mapped epitope. Comparison with available cytoplasmic and flagellar dynein heavy chain sequences reveals regions that are highly conserved in all dynein heavy chains sequenced to date, regions that are conserved only among axonemal dynein heavy chains, and regions that are unique to individual dynein heavy chains. The presumed hydrolytic site is absolutely conserved among dyneins, two other P loops are highly conserved among cytoplasmic dynein heavy chains but not in axonemal dynein heavy chains, and the fourth P loop is invariant in axonemal dynein heavy chains but not in cytoplasmic dynein. One region that is very highly conserved in all dynein heavy chains is similar to a portion of the ATP-sensitive microtubule-binding domain of kinesin. Two other regions present in all dynein heavy chains are predicted to have high alpha-helical content and have a high probability of forming coiled-coil structures. Overall, the central one-third of the gamma dynein heavy chain is most conserved whereas the N-terminal one-third is least conserved; the fact that the latter region is divergent between the cytoplasmic dynein heavy chain and two different axonemal dynein heavy chains suggests that it is involved in chain-specific functions.

scholarly journals Chymotryptic digestion of Tetrahymena ciliary dynein. II. Pathway of the degradation of 22S dynein heavy chains.

1987 ◽  
Vol 105 (2) ◽  
pp. 897-901 ◽  
Author(s):  
Y Y Toyoshima
Keyword(s):  
Atpase Activity ◽  
Gel Electrophoresis ◽  
Peptide Mapping ◽  
Preceding Paper ◽  
Degradation Pathway ◽  
Heavy Chains ◽  
Cell Biol ◽  
Dynein Heavy Chains

As shown in the preceding paper (Toyoshima, Y. Y., 1987, J. Cell Biol., 105:887-895) three-headed Tetrahymena 22S dynein consists of three heavy chains (HCs) and is decomposed into two-headed (H) and one-headed (L) fragments by chymotryptic digestion. To accurately determine the presence of multiple ATPases and ultimately the location of various domains, it is necessary to determine the identity of each HC fragment relative to the original HCs in 22S dynein. The degradation pathway of each HC was determined by peptide mapping and immunoblotting. The three HCs (A alpha, A beta, and A gamma) were immunologically different; although SDS-urea gel electrophoresis showed that A gamma HC was apparently resistant to the digestion, actually three distinct HCs contributed to the same band alternately. H fragment was derived from A beta and A gamma HCs, whereas L fragment originated from A alpha HC. Since both fragments were associated with ATPase activity, these results directly demonstrate the presence of multiple ATPase sites in Tetrahymena 22S dynein.

scholarly journals Chlamydomonas WDR92 in association with R2TP-like complex and multiple DNAAFs to regulate ciliary dynein preassembly

2018 ◽  
Vol 11 (9) ◽  
pp. 770-780 ◽  
Author(s):  
Guang Liu ◽  
Limei Wang ◽  
Junmin Pan
Keyword(s):  
Mass Spectrometry ◽  
Insertional Mutagenesis ◽  
The Other ◽  
Cytoplasmic Protein ◽  
Flagellar Motility ◽  
Heavy Chains ◽  
The Third ◽  
Axonemal Dynein ◽  
Dynein Arms ◽  
Dna Insertional Mutagenesis

Abstract The motility of cilia or eukaryotic flagella is powered by the axonemal dyneins, which are preassembled in the cytoplasm by proteins termed dynein arm assembly factors (DNAAFs) before being transported to and assembled on the ciliary axoneme. Here, we characterize the function of WDR92 in Chlamydomonas. Loss of WDR92, a cytoplasmic protein, in a mutant wdr92 generated by DNA insertional mutagenesis resulted in aflagellate cells or cells with stumpy or short flagella, disappearance of axonemal dynein arms, and diminishment of dynein arm heavy chains in the cytoplasm, suggesting that WDR92 is a DNAAF. Immunoprecipitation of WDR92 followed by mass spectrometry identified inner dynein arm heavy chains and multiple DNAAFs including RuvBL1, RPAP3, MOT48, ODA7, and DYX1C. The PIH1 domain-containing protein MOT48 formed a R2TP-like complex with RuvBL1/2 and RPAP3, while PF13, another PIH1 domain-containing protein with function in dynein preassembly, did not. Interestingly, the third PIH1 domain-containing protein TWI1 was not related to flagellar motility. WDR92 physically interacted with the R2TP-like complex and the other identified DNNAFs. Our data suggest that WDR92 functions in association with the HSP90 co-chaperone R2TP-like complex as well as linking other DNAAFs in dynein preassembly.

scholarly journals The intraflagellar transport dynein complex of trypanosomes is made of a heterodimer of dynein heavy chains and of light and intermediate chains of distinct functions

2014 ◽  
Vol 25 (17) ◽  
pp. 2620-2633 ◽  
Author(s):  
Thierry Blisnick ◽  
Johanna Buisson ◽  
Sabrina Absalon ◽  
Alexandra Marie ◽  
Nadège Cayet ◽  
...  
Keyword(s):  
Protein Complexes ◽  
Intraflagellar Transport ◽  
Retrograde Transport ◽  
High Concentration ◽  
Anterograde Transport ◽  
Heavy Chains ◽  
Cilia And Flagella ◽  
The Stability ◽  
Intermediate Chains ◽  
Dynein Heavy Chains

Cilia and flagella are assembled by intraflagellar transport (IFT) of protein complexes that bring tubulin and other precursors to the incorporation site at their distal tip. Anterograde transport is driven by kinesin, whereas retrograde transport is ensured by a specific dynein. In the protist Trypanosoma brucei, two distinct genes encode fairly different dynein heavy chains (DHCs; ∼40% identity) termed DHC2.1 and DHC2.2, which form a heterodimer and are both essential for retrograde IFT. The stability of each heavy chain relies on the presence of a dynein light intermediate chain (DLI1; also known as XBX-1/D1bLIC). The presence of both heavy chains and of DLI1 at the base of the flagellum depends on the intermediate dynein chain DIC5 (FAP133/WDR34). In the IFT140RNAi mutant, an IFT-A protein essential for retrograde transport, the IFT dynein components are found at high concentration at the flagellar base but fail to penetrate the flagellar compartment. We propose a model by which the IFT dynein particle is assembled in the cytoplasm, reaches the base of the flagellum, and associates with the IFT machinery in a manner dependent on the IFT-A complex.

High molecular weight polypeptides related to dynein heavy chains in Nicotiana tabacum pollen tubes

1995 ◽  
Vol 108 (3) ◽  
pp. 1117-1125
Author(s):  
A. Moscatelli ◽  
C. Del Casino ◽  
L. Lozzi ◽  
G. Cai ◽  
M. Scali ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Nicotiana Tabacum ◽  
Pollen Tube ◽  
High Molecular Weight ◽  
Pollen Tubes ◽  
Biochemical Properties ◽  
Bovine Brain ◽  
Atp Binding ◽  
Heavy Chains ◽  
Dynein Heavy Chains

Nicotiana tabacum pollen tubes contain two high molecular weight polypeptides (about 400 kDa), which are specifically expressed during pollen germination and pollen tube growth in BK medium. The high molecular weight doublet resembles the dynein heavy chains in some biochemical properties. Sedimentation profiles of pollen tube extracts show that the high molecular weight bands have sedimentation coefficients of 22 S and 12 S, respectively. ATPase assay of sedimentation fractions shows an activity ten times higher when stimulated by the presence of bovine brain microtubules in fractions containing the 22 S high molecular weight polypeptide. Both these high molecular weight polypeptides can bind microtubules in an ATP-dependent fashion. A mouse antiserum to a synthetic peptide reproducing the sequence of the most conserved ATP-binding site among dynein heavy chains recognized the two high molecular weight polypeptides. Therefore these polypeptides have sequences immunologically related to the ATP binding sites of dynein heavy chains.

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