Systematic studies of all PIH proteins in zebrafish reveal their distinct roles in axonemal dynein assembly

Yamaguchi, Hiroshi; Oda, Toshiyuki; Kikkawa, Masahide; Takeda, Hiroyuki

doi:10.1101/288837

Cited by 10 publications

(28 citation statements)

References 48 publications

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“…Mutations in DNAAF4/DYX1C/PF23 result in loss of the majority of IDAs and a fraction of ODA in Chlamydomonas flagella (Yamamoto et al, 2017). MOT48 functions in preassembly of IDAs and homologous to the protein interaction with Hsp90 1 (PIH1) domain-containing protein of the R2TP complex (Yamamoto et al, 2010; Yamaguchi et al, 2018). Chlamdyomonas RuvBL1 and RPAP3 are homologs of RuvBL1 and RPAP3 of the human R2TP complex, respectively (Maurizy et al, 2018).…”

Section: Resultsmentioning

confidence: 99%

“…TWI1 is a homolog of PIH1D3, which is implicated dynein arm preassembly in mouse and zebrafish (Dong et al, 2014; Yamaguchi et al, 2018). Currently, it is not known whether TWI1 has a similar function in Chlamydomonas .…”

Section: Resultsmentioning

confidence: 99%

“…R2TP is involved in the assembly of large protein complexes such as RNA polymerase II, small nucleolar ribonucleoproteins (snoRNPs), and protein complexes associated with phosphatidylinositol 3 kinase-related kinases (PIKKs) (Kakihara and Houry, 2012). Recent advances suggest that it also functions in dynein preassembly as revealed by analysis of alteration of genes encoding R2TP components and/or their homologs in various ciliated organisms (Omran et al, 2008; Yamamoto et al, 2010; Knowles et al, 2013; Zhao et al, 2013; Dong et al, 2014; Li et al, 2017; Olcese et al, 2017; Paff et al, 2017; Yamaguchi et al, 2018; Zur Lage et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

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

Liu

Wang

Pan

2018

Journal of Molecular Cell Biology

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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.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Liu

Wang

Pan

2018

Journal of Molecular Cell Biology

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“…Scale bar: 10 nm. ( b ) Combined with genetic perturbation, subtomogram averaging defined the roles of individual dynein assembly factors in the organization of the zebrafish sperm axoneme [ 102 ]. ( c ) Three-dimensional segmentation unveiled the complex organization of the isolated bovine sperm connecting piece, revealing novel fibrous linkers between plates of the segmented columns [ 103 ].…”

Section: Cryo-electron Microscopy Is Poised To Make Important Contribmentioning

confidence: 99%

“…Cryo-ET also provides a means to assess the molecular effects of targeted genetic disruption. Studies in zebrafish revealed how mutations in different dynein assembly factors resulted in structural defects at specific positions in the axoneme ( figure 5 b ), which could be related to altered patterns of sperm motility [ 102 ]. Genetic perturbation can also have larger-scale effects on morphology, and the multi-scale capability of cryo-ET allows it to capture these as well.…”

Section: Cryo-electron Microscopy Is Poised To Make Important Contribmentioning

confidence: 99%

Looking back and looking forward: contributions of electron microscopy to the structural cell biology of gametes and fertilization

2020

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Mammalian gametes—the sperm and the egg—represent opposite extremes of cellular organization and scale. Studying the ultrastructure of gametes is crucial to understanding their interactions, and how to manipulate them in order to either encourage or prevent their union. Here, we survey the prominent electron microscopy (EM) techniques, with an emphasis on considerations for applying them to study mammalian gametes. We review how conventional EM has provided significant insight into gamete ultrastructure, but also how the harsh sample preparation methods required preclude understanding at a truly molecular level. We present recent advancements in cryo-electron tomography that provide an opportunity to image cells in a near-native state and at unprecedented levels of detail. New and emerging cellular EM techniques are poised to rekindle exploration of fundamental questions in mammalian reproduction, especially phenomena that involve complex membrane remodelling and protein reorganization. These methods will also allow novel lines of enquiry into problems of practical significance, such as investigating unexplained causes of human infertility and improving assisted reproductive technologies for biodiversity conservation.

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Ciliary dynein arms: Cytoplasmic preassembly, intraflagellar transport, and axonemal docking

Qiu

Roy

2022

Journal Cellular Physiology

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The microtubular scaffold of motile cilia-the axoneme, is decorated with dynein arms, which are large multiprotein complexes essential for ciliary motility. Dynein arms are arranged along the length of the axoneme in a precise repeating pattern, converting chemical energy from ATP hydrolysis into ciliary mechanical movement.How these complicated molecular machines are assembled coordinately and accurately, starting from mere polypeptide chains in the cytoplasm, remains a fascinating yet perplexing question. Rapidly emerging evidence, from multiple studies carried out with different model organisms and with various methodologies, has highlighted the existence of a dedicated assembly pathway. Here, we summarize recent progress made in clarifying the axonemal dynein arm assembly process, focusing on individual assembly steps, including cytoplasmic preassembly, intraflagellar transport, and axonemal docking. K E Y W O R D S dynein arm assembly, dynein arms, motile cilia 1 | INTRODUCTION Motile cilia are microtubule-based organelles that appear as hair-like protrusions on the cell surface and are present in diverse organisms across many evolutionary lineages. The microtubule scaffold of the cilium, the axoneme, is typically organized in a unique "9 + 2" pattern, in which a pair of central microtubules is surrounded by nine peripheral microtubule doublets (each doublet comprising a complete A and an incomplete B tubule) in a circular arrangement (Figure 1) (Ishikawa et al., 2021; Ishikawa, 2017). Importantly, a variety of biological processes, ranging from the free movement of individual cells such as protozoans like Paramecium and spermatozoa to the flow of mucus over the surface of the respiratory tract and circulation of cerebrospinal fluid within the brain ventricular cavities of mammals, all depend on the coordinated beating of motile cilia (F. Zhou & Roy, 2015).Ciliary motility is imparted by molecular motors known as outer and inner dynein arms (ODAs and IDAs, respectively), which are stably attached to the surface of the A tubule of the peripheral axonemal microtubules (Figure 1) (Ishikawa et al., 2021; Ishikawa, 2017). Dynein arms are very large, complex macromolecular assemblies that consist of multiple protein subunits (the estimated size can range from 1-to 2-MDa), overall contributing for ATP catalysis, microtubule binding activity, as well as regulatory input (King, 2016). The core catalytic subunits are comprised of dynein heavy chains (HCs), which have a length of around 4500 amino acid residues (around 500 KDa in size). Additionally, HCs associate and complex with multiple intermediate chains (ICs) and light chains (LCs), forming the dynein motors. Moreover, with the help of docking complexes (DC), dynein motors are able to stably associate with the axonemal microtubules, thereby enabling sliding motion of adjacent microtubule doublets and producing the overall oscillatory movement of the axoneme.Due to their elaborate structure, absence or mutation of even a single structural component of the dynein...

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Systematic studies of all PIH proteins in zebrafish reveal their distinct roles in axonemal dynein assembly

Cited by 10 publications

References 48 publications

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

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

Looking back and looking forward: contributions of electron microscopy to the structural cell biology of gametes and fertilization

Ciliary dynein arms: Cytoplasmic preassembly, intraflagellar transport, and axonemal docking

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