scholarly journals Cytoplasmic E2f4 forms organizing centres for initiation of centriole amplification during multiciliogenesis

2017 ◽  
Vol 8 (1) ◽  
Author(s):  
Munemasa Mori ◽  
Renin Hazan ◽  
Paul S. Danielian ◽  
John E. Mahoney ◽  
Huijun Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Large Scale ◽  
Abnormal Development ◽  
Basal Bodies ◽  
Transcriptional Program ◽  
Mutant Proteins ◽  
Multiprotein Complexes ◽  
Airway Diseases ◽  
Multiciliated Cells ◽  
Cytoplasmic Structures

Abstract Abnormal development of multiciliated cells is a hallmark of a variety of human conditions associated with chronic airway diseases, hydrocephalus and infertility. Multiciliogenesis requires both activation of a specialized transcriptional program and assembly of cytoplasmic structures for large-scale centriole amplification that generates basal bodies. It remains unclear, however, what mechanism initiates formation of these multiprotein complexes in epithelial progenitors. Here we show that this is triggered by nucleocytoplasmic translocation of the transcription factor E2f4. After inducing a transcriptional program of centriole biogenesis, E2f4 forms apical cytoplasmic organizing centres for assembly and nucleation of deuterosomes. Using genetically altered mice and E2F4 mutant proteins we demonstrate that centriole amplification is crucially dependent on these organizing centres and that, without cytoplasmic E2f4, deuterosomes are not assembled, halting multiciliogenesis. Thus, E2f4 integrates nuclear and previously unsuspected cytoplasmic events of centriole amplification, providing new perspectives for the understanding of normal ciliogenesis, ciliopathies and cancer.

scholarly journals E2F4’s cytoplasmic role in multiciliogenesis is mediated via an N-terminal domain that binds two components of the centriole replication machinery, Deup1 and SAS6

2021 ◽  
pp. mbc.E21-01-0039
Author(s):  
Renin Hazan ◽  
Munemasa Mori ◽  
Paul S. Danielian ◽  
Vincent J. Guen ◽  
Seth M. Rubin ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Reproductive Organs ◽  
Transcriptional Program ◽  
Replication Machinery ◽  
Terminal Domain ◽  
Multiciliated Cells ◽  
N Terminus ◽  
Shared Roles

Multiciliated cells play critical roles in the airway, reproductive organs and brain. Generation of multiple cilia requires both activation of a specialized transcriptional program and subsequent massive amplification of centrioles within the cytoplasm. The E2F4 transcription factor is required for both roles, and consequently for multiciliogenesis. Here, we establish that E2F4 associates with two distinct components of the centriole replication machinery, Deup1 and SAS6, targeting non-homologous domains in these proteins. We map Deup1 and SAS6 binding to E2F4’s N-terminus, and show that this domain is sufficient to mediate E2F4’s cytoplasmic role in multiciliogenesis. This sequence is highly conserved across the E2F family, but the ability to bind Deup1 and SAS6 is specific to E2F4 and E2F5, consistent with their shared roles in multiciliogenesis. By generating E2F4/E2F1 chimeras, we identify a six-residue motif that is critical for Deup1 and SAS6 binding. We propose that the ability of E2F4 and E2F5 to recruit Deup1 and/or SAS6, and enable centriole replication, contributes to their cytoplasmic roles in multiciliogenesis.

scholarly journals The highly conserved FOXJ1 target CFAP161 is dispensable for motile ciliary function in mouse and Xenopus

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Anja Beckers ◽  
Franziska Fuhl ◽  
Tim Ott ◽  
Karsten Boldt ◽  
Magdalena Maria Brislinger ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Cell Surface ◽  
Basal Bodies ◽  
Ciliary Function ◽  
Multiciliated Cells ◽  
Microtubule Motor ◽  
High Sequence Conservation ◽  
Motile Cilia ◽  
Genetic Compensation ◽  
In Cells

AbstractCilia are protrusions of the cell surface and composed of hundreds of proteins many of which are evolutionary and functionally well conserved. In cells assembling motile cilia the expression of numerous ciliary components is under the control of the transcription factor FOXJ1. Here, we analyse the evolutionary conserved FOXJ1 target CFAP161 in Xenopus and mouse. In both species Cfap161 expression correlates with the presence of motile cilia and depends on FOXJ1. Tagged CFAP161 localises to the basal bodies of multiciliated cells of the Xenopus larval epidermis, and in mice CFAP161 protein localises to the axoneme. Surprisingly, disruption of the Cfap161 gene in both species did not lead to motile cilia-related phenotypes, which contrasts with the conserved expression in cells carrying motile cilia and high sequence conservation. In mice mutation of Cfap161 stabilised the mutant mRNA making genetic compensation triggered by mRNA decay unlikely. However, genes related to microtubules and cilia, microtubule motor activity and inner dyneins were dysregulated, which might buffer the Cfap161 mutation.

scholarly journals Multimorbidity clusters in patients with chronic obstructive airway diseases in the EpiChron Cohort

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jonás Carmona-Pírez ◽  
Beatriz Poblador-Plou ◽  
Ignatios Ioakeim-Skoufa ◽  
Francisca González-Rubio ◽  
Luis Andrés Gimeno-Feliú ◽  
...  
Keyword(s):  
Substance Use ◽  
Chronic Conditions ◽  
Large Scale ◽  
Mental Health Problems ◽  
Epidemiological Studies ◽  
Chronic Obstructive ◽  
High Morbidity ◽  
Airway Diseases ◽  
Obstructive Sleep ◽  
Obstructive Airway Diseases

AbstractChronic obstructive airway diseases such as chronic obstructive pulmonary disease (COPD), asthma, rhinitis, and obstructive sleep apnea (OSA) are amongst the most common treatable and preventable chronic conditions with high morbidity burden and mortality risk. We aimed to explore the existence of multimorbidity clusters in patients with such diseases and to estimate their prevalence and impact on mortality. We conducted an observational retrospective study in the EpiChron Cohort (Aragon, Spain), selecting all patients with a diagnosis of allergic rhinitis, asthma, COPD, and/or OSA. The study population was stratified by age (i.e., 15–44, 45–64, and ≥ 65 years) and gender. We performed cluster analysis, including all chronic conditions recorded in primary care electronic health records and hospital discharge reports. More than 75% of the patients had multimorbidity (co-existence of two or more chronic conditions). We identified associations of dermatologic diseases with musculoskeletal disorders and anxiety, cardiometabolic diseases with mental health problems, and substance use disorders with neurologic diseases and neoplasms, amongst others. The number and complexity of the multimorbidity clusters increased with age in both genders. The cluster with the highest likelihood of mortality was identified in men aged 45 to 64 years and included associations between substance use disorder, neurologic conditions, and cancer. Large-scale epidemiological studies like ours could be useful when planning healthcare interventions targeting patients with chronic obstructive airway diseases and multimorbidity.

scholarly journals Multiciliated cell basal bodies align in stereotypical patterns coordinated by the apical cytoskeleton

2016 ◽  
Vol 214 (5) ◽  
pp. 571-586 ◽  
Author(s):  
Elisa Herawati ◽  
Daisuke Taniguchi ◽  
Hatsuho Kanoh ◽  
Kazuhiro Tateishi ◽  
Shuji Ishihara ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Imaging System ◽  
Basal Bodies ◽  
Large Numbers ◽  
Multiciliated Cells ◽  
Alignment Process ◽  
Flow Through ◽  
Linear Alignment ◽  
Green Fluorescent

Multiciliated cells (MCCs) promote fluid flow through coordinated ciliary beating, which requires properly organized basal bodies (BBs). Airway MCCs have large numbers of BBs, which are uniformly oriented and, as we show here, align linearly. The mechanism for BB alignment is unexplored. To study this mechanism, we developed a long-term and high-resolution live-imaging system and used it to observe green fluorescent protein–centrin2–labeled BBs in cultured mouse tracheal MCCs. During MCC differentiation, the BB array adopted four stereotypical patterns, from a clustering “floret” pattern to the linear “alignment.” This alignment process was correlated with BB orientations, revealed by double immunostaining for BBs and their asymmetrically associated basal feet (BF). The BB alignment was disrupted by disturbing apical microtubules with nocodazole and by a BF-depleting Odf2 mutation. We constructed a theoretical model, which indicated that the apical cytoskeleton, acting like a viscoelastic fluid, provides a self-organizing mechanism in tracheal MCCs to align BBs linearly for mucociliary transport.

scholarly journals Histone demethylase JMJD2B/KDM4B regulates transcriptional program via distinctive epigenetic targets and protein interactors for the maintenance of trophoblast stem cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kylie Hin-Man Mak ◽  
Yuk Man Lam ◽  
Ray Kit Ng
Keyword(s):  
Stem Cells ◽  
Transcription Factor ◽  
Cell Fate ◽  
Histone Demethylase ◽  
Gene Promoters ◽  
Transcriptional Program ◽  
Binding Motifs ◽  
Trophoblast Stem Cells ◽  
Trophoblast Stem ◽  
Trophoblast Stem Cell

AbstractTrophoblast stem cell (TSC) is crucial to the formation of placenta in mammals. Histone demethylase JMJD2 (also known as KDM4) family proteins have been previously shown to support self-renewal and differentiation of stem cells. However, their roles in the context of the trophoblast lineage remain unclear. Here, we find that knockdown of Jmjd2b resulted in differentiation of TSCs, suggesting an indispensable role of JMJD2B/KDM4B in maintaining the stemness. Through the integration of transcriptome and ChIP-seq profiling data, we show that JMJD2B is associated with a loss of H3K36me3 in a subset of embryonic lineage genes which are marked by H3K9me3 for stable repression. By characterizing the JMJD2B binding motifs and other transcription factor binding datasets, we discover that JMJD2B forms a protein complex with AP-2 family transcription factor TFAP2C and histone demethylase LSD1. The JMJD2B–TFAP2C–LSD1 complex predominantly occupies active gene promoters, whereas the TFAP2C–LSD1 complex is located at putative enhancers, suggesting that these proteins mediate enhancer–promoter interaction for gene regulation. We conclude that JMJD2B is vital to the TSC transcriptional program and safeguards the trophoblast cell fate via distinctive protein interactors and epigenetic targets.

Genetic analysis of pattern formation in the Arabidopsis embryo

Development ◽  
1991 ◽  
Vol 113 (Supplement_1) ◽  
pp. 27-38 ◽  
Author(s):  
Gerd Jürgens ◽  
Ulrike Mayer ◽  
Torres Ruiz Ramon A. ◽  
Thomas Berleth ◽  
Simon Miséra
Keyword(s):  
Pattern Formation ◽  
Large Scale ◽  
Early Embryo ◽  
Flowering Plant ◽  
Abnormal Development ◽  
Mutant Seedling ◽  
Basic Body ◽  
Plant Embryo ◽  
Embryonic Pattern Formation ◽  
Plant Arabidopsis Thaliana

Virtually nothing is known about the mechanisms that generate the basic body pattern in plant embryogenesis. As a first step towards the analysis of pattern formation, we have isolated and begun to characterise putative pattern mutants in the flowering plant, Arabidopsis thaliana. A large-scale screen for morphologically abnormal seedling mutants yielded about 250 lines for further study, and genetic evidence suggests saturation of the genome for this kind of mutation. The phenotypes of putative pattern mutants fall into distinct categories, classes and groups, which may reflect specific aspects of embryonic pattern formation. Mutant seedling phenotypes result from abnormal development in the early embryo. The implications of our findings are discussed with regard to the prospects for a mechanistic understanding of pattern formation in the plant embryo.

Isolation and characterization of a downstream target of Pax6 in the mammalian retinal primordium

Development ◽  
2001 ◽  
Vol 128 (20) ◽  
pp. 3987-3994 ◽  
Author(s):  
Gilbert Bernier ◽  
Wolfgang Vukovich ◽  
Lorenz Neidhardt ◽  
Bernhard G. Herrmann ◽  
Peter Gruss
Keyword(s):  
Transcription Factor ◽  
Expression Pattern ◽  
Large Scale ◽  
Ectopic Expression ◽  
Signal Transduction Pathway ◽  
Homeobox Gene ◽  
Optic Vesicle ◽  
Altered Expression ◽  
Retina Development ◽  
Isolation And Characterization

The transcription factor Pax6 is required for eye morphogenesis in humans, mice and insects, and can induce ectopic eye formation in vertebrate and invertebrate organisms. Although the role of Pax6 has intensively been studied, only a limited number of genes have been identified that depend on Pax6 activity for their expression in the mammalian visual system. Using a large-scale in situ hybridization screen approach, we have identified a novel gene expressed in the mouse optic vesicle. This gene, Necab, encodes a putative cytoplasmic Ca2+-binding protein and coincides with Pax6 expression pattern in the neural ectoderm of the optic vesicle and in the forebrain pretectum. Remarkably, Necab expression is absent in both structures in Pax6 mutant embryos. By contrast, the optic vesicle-expressed homeobox genes Rx, Six3, Otx2 and Lhx2 do not exhibit an altered expression pattern. Using gain-of-function experiments, we show that Pax6 can induce ectopic expression of Necab, suggesting that Necab is a direct or indirect transcriptional target of Pax6. In addition, we have found that Necab misexpression can induce ectopic expression of the homeobox gene Chx10, a transcription factor implicated in retina development. Taken together, our results provide evidence that Necab is genetically downstream of Pax6 and that it is a part of a signal transduction pathway in retina development.

scholarly journals Celsr1 and CAMSAP3 differently regulate intercellular and intracellular cilia orientation in oviduct multiciliated cells

2020 ◽  
Author(s):  
Fumiko Matsukawa Usami ◽  
Masaki Arata ◽  
Dongbo Shi ◽  
Sanae Oka ◽  
Yoko Higuchi ◽  
...  
Keyword(s):  
Cell Polarity ◽  
Planar Cell Polarity ◽  
Molecular Mechanisms ◽  
The Other ◽  
Basal Bodies ◽  
Generating Functional ◽  
Tissue Polarity ◽  
Other Hand ◽  
Multiciliated Cells ◽  
Polarity Regulation

SummaryThe molecular mechanisms by which cilia orientation is coordinated within and between multiciliated cells (MCCs) is not fully understood. By observing the orientation of basal bodies (BB) in MCCs of mouse oviducts, here, we show that Celsr1, a planar cell polarity (PCP) factor involved in tissue polarity regulation, is dispensable for determining BB orientation in individual cells, whereas CAMSAP3, a microtubule minus-end regulator, is critical for this process but not for PCP. MCCs exhibit a characteristic BB orientation and microtubule gradient along the tissue axis, and these intracellular polarities were maintained in the cells lacking Celsr1, although the intercellular coordination of the polarities was partly disrupted. On the other hand, CAMSAP3 regulated the assembly of microtubules interconnecting BBs by localizing at the BBs, and its mutation led to disruption of intracellular coordination of BB orientation, but not affecting PCP factor localization. Thus, both Celsr1 and CAMSAP3 are responsible for BB orientation but in distinct ways; and therefore, their cooperation should be critical for generating functional multiciliated tissues.

Mutations affecting development of the zebrafish inner ear and lateral line

Development ◽  
1996 ◽  
Vol 123 (1) ◽  
pp. 241-254 ◽  
Author(s):  
T.T. Whitfield ◽  
M. Granato ◽  
F.J. van Eeden ◽  
U. Schach ◽  
M. Brand ◽  
...  
Keyword(s):  
Inner Ear ◽  
Lateral Line ◽  
Large Scale ◽  
Sensory System ◽  
Semicircular Canals ◽  
Abnormal Development ◽  
Pigment Cells ◽  
Otic Vesicle ◽  
In The Wild ◽  
Complementation Groups

Mutations giving rise to anatomical defects in the inner ear have been isolated in a large scale screen for mutations causing visible abnormalities in the zebrafish embryo (Haffter, P., Granato, M., Brand, M. et al. (1996) Development 123, 1–36). 58 mutants have been classified as having a primary ear phenotype; these fall into several phenotypic classes, affecting presence or size of the otoliths, size and shape of the otic vesicle and formation of the semicircular canals, and define at least 20 complementation groups. Mutations in seven genes cause loss of one or both otoliths, but do not appear to affect development of other structures within the ear. Mutations in seven genes affect morphology and patterning of the inner ear epithelium, including formation of the semicircular canals and, in some, development of sensory patches (maculae and cristae). Within this class, dog-eared mutants show abnormal development of semicircular canals and lack cristae within the ear, while in van gogh, semicircular canals fail to form altogether, resulting in a tiny otic vesicle containing a single sensory patch. Both these mutants show defects in the expression of homeobox genes within the otic vesicle. In a further class of mutants, ear size is affected while patterning appears to be relatively normal; mutations in three genes cause expansion of the otic vesicle, while in little ears and microtic, the ear is abnormally small, but still contains all five sensory patches, as in the wild type. Many of the ear and otolith mutants show an expected behavioural phenotype: embryos fail to balance correctly, and may swim on their sides, upside down, or in circles. Several mutants with similar balance defects have also been isolated that have no obvious structural ear defect, but that may include mutants with vestibular dysfunction of the inner ear (Granato, M., van Eeden, F. J. M., Schach, U. et al. (1996) Development, 123, 399–413,). Mutations in 19 genes causing primary defects in other structures also show an ear defect. In particular, ear phenotypes are often found in conjunction with defects of neural crest derivatives (pigment cells and/or cartilaginous elements of the jaw). At least one mutant, dog-eared, shows defects in both the ear and another placodally derived sensory system, the lateral line, while hypersensitive mutants have additional trunk lateral line organs.

Sign in / Sign up

Export Citation Format

Share Document