A Genome-Wide RNAi Screen to Dissect Centriole Duplication and Centrosome Maturation in Drosophila

Dobbelaere, Jeroen; Josué, Filipe; Suijkerbuijk, Saskia J.E.; Baum, Buzz; Tapon, Nicolas; Raff, Jordan W.

doi:10.1371/journal.pbio.0060224

Cited by 224 publications

(287 citation statements)

References 72 publications

Supporting

Mentioning

266

Contrasting

Unclassified

Order By: Relevance

“…In Drosophila S2Rϩ cells, RNAi knockdown of Bld10 gave an intermediate centriole replication phenotype (Dobbelaere et al, 2008). This contrast with our in vivo results, likely reflects a peculiarity of S2Rϩ cells or possible off-target effects of RNAi.…”

Section: Proper Assembly Of Centrioles and Basal Bodies Require Bld10contrasting

confidence: 99%

Section: Bld10 Is a Centriolar And Sperm Basal Body Proteinsupporting

confidence: 53%

“…These data indicate that Bld10 is likely a ubiquitous component of centrioles throughout development. Two recent reports corroborated the centriole localization of Bld10 (Blachon et al, 2008;Dobbelaere et al, 2008).In Drosophila spermatocytes, where the centrioles are ϳ10-fold longer than in somatic cells (Gonzalez et al, 1998), the centriole-specific marker YFP-Asl (Varmark et al, 2007) colocalized with Bld10 at centrioles throughout spermatocyte development. In mature spermatocytes and throughout the two meiotic divisions, Bld10 was concentrated at the proximal end of the centrioles, a subregion of the centriole where YFP-Asl and Bld10 localization overlapped ( Figure 2B).…”

mentioning

confidence: 73%

See 2 more Smart Citations

DrosophilaBld10 Is a Centriolar Protein That Regulates Centriole, Basal Body, and Motile Cilium Assembly

Mottier-Pavie

Megraw

2009

MBoC

107

View full text Add to dashboard Cite

Cilia and flagella play multiple essential roles in animal development and cell physiology. Defective cilium assembly or motility represents the etiological basis for a growing number of human diseases. Therefore, how cilia and flagella assemble and the processes that drive motility are essential for understanding these diseases. Here we show that Drosophila Bld10, the ortholog of Chlamydomonas reinhardtii Bld10p and human Cep135, is a ubiquitous centriolar protein that also localizes to the spermatid basal body. Mutants that lack Bld10 assemble centrioles and form functional centrosomes, but centrioles and spermatid basal bodies are short in length. bld10 mutant flies are viable but male sterile, producing immotile sperm whose axonemes are deficient in the central pair of microtubules. These results show that Drosophila Bld10 is required for centriole and axoneme assembly to confer cilium motility. INTRODUCTIONCentrioles lie at the core of centrosomes. They consist of a ninefold symmetrical array of nine triplet microtubules arranged in a cylinder. In most differentiated cell types, centrioles transform into basal bodies, membrane-embedded centrioles that template cilium and flagellum axoneme assembly. The requirement of cilia and flagella for many developmental and physiological processes, together with the growing list of human diseases that result from defects in basal bodies and cilia (Badano et al., 2006;Bisgrove and Yost, 2006;Bettencourt-Dias and Glover, 2007;Fliegauf et al., 2007;Marshall, 2008), drives the need to understand the basic processes involved in centriole, basal body, cilium assembly, and motility.In Drosophila several approaches have identified evolutionarily conserved centriole proteins required for centriole biogenesis including Sak, Sas4, Sas6, Ana1, Ana2, and Asterless (Bettencourt-Dias et al., 2005;Basto et al., 2006;Goshima et al., 2007;Rodrigues-Martins et al., 2007;Blachon et al., 2008). In flies, mutations in these genes abolish centrosome and cilium/flagellum assembly (except mutations in ana1 and ana2, which have not been described yet). Additional centriole/ basal body proteins including Spd-2, Drosophila pericentrin-like protein (D-PLP)/CP309, and uncoordinated (UNC) function in pericentriolar material (PCM) recruitment to centrosomes and/or the assembly of cilia and flagella Kawaguchi and Zheng, 2004;Martinez-Campos et al., 2004;Dix and Raff, 2007;Giansanti et al., 2008). Identification of the complete set of centriole components is necessary to define their individual and cooperative roles in the assembly and function of centrioles and cilia.In Chlamydomonas reinhardtii, mutations in the bld10 gene result in complete loss of flagella, giving the cells a "bald" appearance, due to a failure to assemble centrioles (Matsuura et al., 2004;Hiraki et al., 2007). Bld10p functions in the formation of the cartwheel, a ninefold symmetrical scaffold structure essential for an early step of centriole/basal body assembly . From RNA interference (RNAi) studies in cell culture, the human ort...

show abstract

Section: Proper Assembly Of Centrioles and Basal Bodies Require Bld10contrasting

confidence: 99%

Section: Bld10 Is a Centriolar And Sperm Basal Body Proteinsupporting

confidence: 53%

mentioning

confidence: 73%

See 1 more Smart Citation

DrosophilaBld10 Is a Centriolar Protein That Regulates Centriole, Basal Body, and Motile Cilium Assembly

Mottier-Pavie

Megraw

2009

MBoC

107

View full text Add to dashboard Cite

show abstract

“…Many proteins are involved in the regulation of centrosome maturation 10 and, importantly, disruption of these proteins leads to defects in spindle formation similar to the defects seen with Kin-1 depletion [14][15][16] . Such proteins are collectively known as PCM proteins and include Plk-1, Cdk5rap2 and Aik-A.…”

Section: Resultsmentioning

confidence: 99%

Kindlin-1 regulates mitotic spindle formation by interacting with integrins and Plk-1

et al. 2013

View full text Add to dashboard Cite

Kindlin-1 binds to integrins and regulates integrin activation at cell adhesions. Here we report a new function of Kindlin-1 in regulating spindle assembly. We show that Kindlin-1 localizes to centrosomes, its concentration peaking during G2/M, where it associates with various pericentriolar material proteins, including Polo-like kinase 1. Short interfering RNA-mediated depletion of Kindlin-1 increases formation of abnormal mitotic spindles and decreases cellular survival. This effect is dependent not only on the ability of Kindlin-1 to bind integrins but also on Polo-like kinase 1-mediated Kindlin-1 phosphorylation. We demonstrate that a subcellular pool of phosphorylated Kindlin-1 is located exclusively at centrosomes. Our work identifies a novel cellular role for Kindlin-1 in ensuring mitotic spindle assembly and cellular survival that is controlled by phosphorylation via Polo-like kinase 1.

show abstract

“…In flies, where the pathway of centrosome maturation has been more extensively studied, the Pericentrin-like protein (D-PLP) shares this role with centrosomin (Cnn; Martinez-Campos et al 2004). The localisation of both these PCM proteins is dependent upon the presence of the core centriolar component D-Spd2 (Dix and Raff 2007;Giansanti et al 2008), whilst the phosphorylation of Cnn by the mitotic kinase, Polo, correlates with centrosome maturation (Dobbelaere et al 2008), suggesting a hierarchical pathway in which γ-tubulin is targeted and maintained at the PCM in a stepwise fashion.…”

Section: Centrosome-nucleated Mtsmentioning

confidence: 99%

50 ways to build a spindle: the complexity of microtubule generation during mitosis

Duncan

Wakefield

2011

Chromosome Res

View full text Add to dashboard Cite

The accurate segregation of duplicated chromosomes, essential for the development and viability of a eukaryotic organism, requires the formation of a robust microtubule (MT)-based spindle apparatus. Entry into mitosis or meiosis precipitates a cascade of signalling events which result in the activation of pathways responsible for a dramatic reorganisation of the MT cytoskeleton: through changes in the properties of MT-associated proteins, local concentrations of free tubulin dimer and through enhanced MT nucleation. The latter is generally thought to be driven by localisation and activation of γ-tubulin-containing complexes (γ-TuSC and γ-TuRC) at specific subcellular locations. For example, upon entering mitosis, animal cells concentrate γ-tubulin at centrosomes to tenfold the normal level during interphase, resulting in an aster-driven search and capture of chromosomes and bipolar mitotic spindle formation. Thus, in these cells, centrosomes have traditionally been perceived as the primary microtubule organising centre during spindle formation. However, studies in meiotic cells, plants and cell-free extracts have revealed the existence of complementary mechanisms of spindle formation, mitotic chromatin, kinetochores and nucleation from existing MTs or the cytoplasm can all contribute to a bipolar spindle apparatus. Here, we outline the individual known mechanisms responsible for spindle formation and formulate ideas regarding the relationship between them in assembling a functional spindle apparatus.

show abstract

A Genome-Wide RNAi Screen to Dissect Centriole Duplication and Centrosome Maturation in Drosophila

Cited by 224 publications

References 72 publications

DrosophilaBld10 Is a Centriolar Protein That Regulates Centriole, Basal Body, and Motile Cilium Assembly

DrosophilaBld10 Is a Centriolar Protein That Regulates Centriole, Basal Body, and Motile Cilium Assembly

Kindlin-1 regulates mitotic spindle formation by interacting with integrins and Plk-1

50 ways to build a spindle: the complexity of microtubule generation during mitosis

Contact Info

Product

Resources

About