Cell Cycle Regulation

Research Interests

Our research focuses on cell cycle progression and the cytoskeleton in normal development and disease. We are particularly interested in the role played by microtubule organizing structures, such as the centrosome, cilia and flagella. The centrosome is the major microtubule organizer in animal cells, and is very often abnormal in cancer. Cilia and flagella are cellular projections which are indispensable in a variety of cellular and developmental processes including cell motility, propagation of morphogenic signals and sensory reception. Despite their importance, we know very little about centrosome and cilia biogenesis or how they may go awry in human disease. Our laboratory uses an integrated approach to study those questions: we combine studies in model organisms with studies in human cells, bioinformatics and mathematical modeling to have an integrated view of this process. The fruit fly is an excellent organism to address those questions, since it combines possibilities of screening multiple genes with the ability to perform in-depth regulation studies in the whole organism. As the regulatory mechanisms of the cell cycle and cytoskeleton have been highly conserved throughout evolution, we can extrapolate our findings to humans to test their relevance for human disease. An understanding of the pathways involved in cell cycle and cytoskeleton can generate diagnostic and prognostic markers and hopefully provide novel therapeutic targets in human disease.

Check our website at http://sites.igc.gulbenkian.pt/ccr/

Check for opportunities in our group at http://sites.igc.gulbenkian.pt/ccr/opportunities.html

Group Members

Carla Lopes	Postdoc
	Tel: 21 440 7925
Daniela Brito	Postdoc
	Tel: 21 446 4636
Adan Guerrero	Postdoc
	Tel: 21 440 7945
Mariana Faria	Postdoc
	Tel: 21 446 4636
Susana Gouveia	Postdoc
	Tel: 21 440 7925
Gaelle Marteil	Postdoc
	Tel: 21 440 7925
Swadhin Jana	Postdoc
	Tel: 21 440 7970
Marta Mesquita	External Ph.D. Student
	Tel: 21 440 7925
Filipe Leal	External Ph.D. Student
	Tel: 21 440 7925
Pedro Machado	Trainee
	Tel: 21 440 7925
Paulo Duarte	Trainee
	Tel: 21 440 7925
Tiago Amado	Trainee
	Tel: 21 440 7925
Samuel Gilberto	Trainee
	Tel: 21 440 7945
Sónia Rosa	Trainee
	Tel: 21 446 4636
Zita Santos	2006 PDIGC PhD Student
	Tel: 21 440 7925
Inês Ferreira	2007 PGD PhD Student
	Tel: 21 446 4514
Inês Bento	2008 PGD PhD Student
	Tel: 21 440 7925

Research Project

Control of Centriole Structure And Number

Centrioles are essential for the formation of several microtubule organizing structures including cilia, flagella and centrosomes. These structures are involved in a variety of functions, from cell motility to division. Centrosome defects are seen in many cancers, while abnormalities in cilia and flagella can lead to a variety of human diseases, such as polycystic kidney disease. The molecular mechanisms regulating centriole biogenesis have only recently started to be unravelled, opening new ways to answer a wide range of questions that have fascinated biologists for more than a century. In this grant we are asking two fundamental questions that are central to human disease: how is centriole structure and number established and regulated in the eukaryotic cell? To address these questions we propose to identify new molecular players, and to test the role of these and known players in the context of specific mechanistic hypothesis, using in vitro and in vivo models. We propose to develop novel assays for centriole structure and regulation in order to address mechanistic problems not accessible with today’s assays. In our search for novel components we will use a multidisciplinary approach combining bioinformatics with high throughput screening. The use of in vitro systems will permit the quantitative dissection of molecular mechanisms, while the study of those mechanisms in Drosophila will allow us to understand them at the whole organism level. Furthermore, this analysis, together with studies in human tissue culture cells, will allow us to understand the consequences of misregulation of these fundamental centriole properties for human disease, such as ciliopathies and cancer. My group is already collaborating with medical doctors in the study of centriole aberrations in human disease (cancer and ciliopathies), which will be invaluable to bringing the results of this study to the translational level.

Funding

2011-2015- European Research Council-Starting Grant-

Research Project

Role of Centrosome and Ploidy Changes in Tumor progression (Barrett Esophagus)

Barrett’s esophagus is a clinically important premalignant condition. It is a precursor of esophagus adenocarcinoma, a tumor whose incidence has increased over the last decades at a rate exceeding that of any other cancer type. Barrett´s esophagus (BE) has been extensively studied, being particularly important in revealing the prevalence of polyploidization - doubling of the number of chromosome sets - early in tumorigenesis. Despite the known association of BE with polyploidy and its use as a risk factor, a key unstudied issue in the pathogenesis of BE is the potential relationship between centrosome abnormalities and cancer initiation and progression. BE and BAA are unique in that progressive stages of tumor development can be studied because of the availability of serial biopsies from the same patient. The experiments to be performed in this project aim to exploit clinical resources available for BE together with in vitro approaches and animal models to develop what will be a unique collaborative program to attack the broadly important problem of how centrosome abnormalities contribute to tumorigenesis.

Funding

2010-2013-Harvard Medical School-FCT Programme (funded by the Portuguese Agency for Funding Science and Technology-FCT)

Collaborators

Collaborative grant with the laboratories of José Pereira-Leal (Computational Genomics Laboratory @ IGC); Paula Chaves (Pathology laboratory @ Lisbon Cancer Institute- IPO) ; David Pellman (Danna Farber- Harvard Medical School) ; Max Loda (Pathology Laboratory- Danna Farber- Harvard Medical School).

Research Project

Regulation of Cilia and Flagella Biogenesis

Cilia are cellular projections involved in a variety of processes from cell motility to fluid flow and sensing mechanical stimuli. Defects in cilia can cause multiple human diseases, including obesity, retinal degeneration, and male infertility. It is thus of paramount importance to understand the biogenesis of those structures. The core skeleton of cilia, the axoneme, is templated by the centriole/basal body, a structure that tethers at the cytoplasmic membrane. Centrioles are cylinders of nine microtubule (MT) triplets with a nine-fold symmetry, which is provided by a structure called cartwheel. Centrioles are also essential for the formation of centrosomes, the primary MT organiser in the cell. This duality of centriole/basal body function, as part of the centrosome or as a cilium nucleator, is inbuilt into its cellular role. In this project we focus on the centriole to basal body differentiation, at which point centrioles assemble a distal area called Transition Zone (TZ) that is essential for cilia formation. It is in the TZ that growth of the most external MT in the centriole triplets stops, giving rise to the axoneme structure with nine doublets. Moreover, in motile cilia, the TZ assembles a central pair (CP) of MTs, essential for motility of most cilia. Inhibition of TZ formation in fruit flies leads to uncoordination and male sterility, a consequence of defects in sensory neurons and spermatozoa, the two ciliated cell types in the fly.

Funding

2010-2013 PTDC/BIA-BCM/105602/2008- FCT

Collaborators

Giuliano Callaini, University of Siena, Italy.

Research Project

Cellular and Developmental Mechanisms Controlling Centrosome Number

Centrosomes are the major microtubule organising center (MTOC) in animal cells. Most proliferating cells have one centrosome, which duplicates in S phase in coordination with the chromosome cycle. In mitosis the centrosomes help to form the poles of the spindle, each centrosome is then inherited by one daughter cell. Exceptions to that cycle are found in Nature, both during embryonic development and in human disease. For example, centrosomes are eliminated during oogenesis in many animal species; it is the sperm that brings the first centriole to the embryo upon fertilization. Control of centrosome elimination and formation is indispensable during oogenesis, as injection of these organelles is sufficient to start parthenogesis in Xenopus. De novo centrosome formation also occurs naturally in eggs of parthenogenic species. Many cancers have supranumerary and aberrant centrosomes, and these abnormalities correlate with tumour progression. This project aims to understand how centrosome number is controlled, in particular the coordination of the chromosome and canonical centrosome cycles and the regulation of de novo formation. We are using both the Xenopus cell-free egg extract, which has been the biochemical system of excellence to study the cell cycle and cytoskeleton, and Drosophila melanogaster, which provides us with the advantage of 100 years of genetics, rapid RNAi screening assays and cell biology tools to assay centrosome biogenesis in development. This research will lead us to a better understanding of centriole biogenesis, how it is normally regulated in proliferating cells and during critical stages of development, such as oogenesis. Moreover, it may provide us with insight into centrosome number regulation in cancer.

Funding

2010-2013 PTDC/SAU-OBD/105616/2008- FCT

Collaborators

Eric Karsenti & Steffi Kandel Lewis (EMBL)

Research Project

Regulation of Centriole Biogenesis and Function: an Integrative Approach

Using computational and experimental studies, in collaboration with the Computational Genomics Laboratory at IGC, we proposed that a conserved group of proteins explains the conservation of centriole architecture, and that taxon and tissue-specific molecular innovations, gained through emergence, or duplication and divergence, play important roles in coordinating centriole biogenesis to different cellular contexts. This work contextualises the biogenesis and functions of the centriole structure, providing us with a conceptual, evolutionary framework, and suggesting strategies that we are now using to identify novel players in those processes (Carvalho-Santos et al, JCS, 2010). With the Computational Genomics laboratory, we are creating phylogenetic profiles of molecular players in centriole assembly and correlating them with the evolution of the morphology of centrioles using mostly published electron micrographs. We are assembling a database using controlled vocabulary and linking protein repertoires with morphological diversity (www.centrioledb.org). This database is built by a consortium of International experts in the biology of each organisms, including as of September 2010, more than 25 laboratories in Europe, the Americas and Japan. These (taxonomic-)domain experts upload Electron Microscopy images accompanied by detailed structural annotations by means of a controlled vocabulary developed specifically for this task. The collection of these images is supplemented by manually curated lists of proteins known to be involved in the assembly of centrioles and their related structures. Finally, automatic orthology mapping is used to generate a phylogenomic profile, allowing direct comparison between molecular data and morphological differences in an evolutionary context. Currently centrioleDB consists of hundreds of images from over 70 species, and genomes from nearly 250 species. As the knowledge-base expands it will provide a powerful tool for cell biologists to predict biologically relevant relationships between proteins and mtoc structure and function, as well as providing insights to the evolutionary mechanisms involved in creating structural variation. We anticipate that this database will catalyze a new dimension of cell biological research.

Funding

PTDC/BIA-BCM/73195/2006, FCT

Collaborators

José Pereira Leal (IGC)
Juliette Azimzadeh (UCSF)
Michel Bornens (Curie Institute)
Keith Gull (University of Oxford)
More than 25 laboratories in Europe, the Americas and Japan.

Research Project

Regulation of Centriole Assembly

The centrosome is the primary microtubule organizing centre (MTOC) in animal cells. It includes two centrioles, which act as a core structure and can also function as basal bodies to grow flagella or cilia. Recent studies, including ours, have identified a group of conserved proteins which give us an entry point to those questions. The assembly programme starts with recruitment of SAK, a kinase involved in tumorigenesis, to the mother centriole. This allows recruitment of SAS-6 and SAS-4, two coiled-coil molecules necessary for the formation of a “centriole scaffold” and recruitment of centriolar MTs, respectively. How those molecules organise microtubules to form a centriole is not known. Here we will search for molecular interactors of those proteins and explore novel assays in centriole biogenesis.

Funding

EMBO Installation Grant, European Molecular Biology Organisation (EMBO), Fundação para a Ciência e a Tecnologia (FCT, Portugal) and Fundação Calouste

Collaborators

CRMB, Montpellier, France - Carsten Janke
EMBL, Heidelberg, Germany - Eric Karsenti
Department of Genetics, University of Cambridge, UK - David Glover

Publications

(selected) Updated April (2010).

NS, Yu QD, Weiskopf K, Tzolovsky G, Cunha-Ferreira I, Riparbelli M, Rodrigues-Martins A, Bettencourt Dias M, Callaini G, Glover DM (2010). Asterless is a scaffold for the onset of centriole assembly. Dzhindzhev Nature [Epub ahead of print]

Otto EA, Hurd TW, Airik R, Chaki M, Zhou W, Stoetzel C, Patil SB, Levy S, Ghosh AK, Murga-Zamalloa CA, van Reeuwijk J, Letteboer SJ, Sang L, Giles RH, Liu Q, Coene KL, Estrada-Cuzcano A, Collin RW, McLaughlin HM, Held S, Kasanuki JM, Ramaswami G, Conte J, Lopez I, Washburn J, Macdonald J, Hu J, Yamashita Y, Maher ER, Guay-Woodford LM, Neumann HP, Obermüller N, Koenekoop RK, Bergmann C, Bei X, Lewis RA, Katsanis N, Lopes V, Williams DS, Lyons RH, Dang CV, Brito DA, Bettencourt Dias M, Zhang X, Cavalcoli JD, Nürnberg G, Nürnberg P, Pierce EA, Jackson PK, Antignac C, Saunier S, Roepman R, Dollfus H, Khanna H, Hildebrandt F. (2010). Candidate exome capture identifies mutation of SDCCAG8 as the cause of a retinal-renal ciliopathy. Nat Genet [Epub ahead of print]

Mónica Bettencourt Dias (2010). Centered on centrioles. Interview by Ben Short. J Cell Biol 190(5) :710-1

Martins AR, Machado P, Callaini G, Bettencourt Dias M. (2010). Microscopy methods for the study of centriole biogenesis and function in Drosophila. Methods Cell Biol 97 :223-42

Carvalho-Santos Z, Machado P, Branco P, Tavares-Cadete F, Rodrigues-Martins A, Pereira-Leal JB, Bettencourt Dias, M.() (2010). Stepwise evolution of the centriole-assembly pathway J Cell Sci.

Debec A, Sullivan W, Bettencourt Dias, M. (2010). Centrioles: active players or passengers during mitosis? Cell Mol Life Sci

Kuriyama R, Bettencourt Dias, M, Hoffmann I, Arnold M, Sandvig L. (2009). Gamma-tubulin-containing abnormal centrioles are induced by insufficient Plk4 in human HCT116 colorectal cancer cells J Cell Sci. 15;122(Pt 12) :2014-23

Cunha-Ferreira I, Rodrigues-Martins A, Bento I, Riparbelli M, Zhang W, Laue E, Callaini G, Glover DM, Bettencourt Dias M. (2009). The SCF/Slimb Ubiquitin Ligase Limits Centrosome Amplification through Degradation of SAK/PLK4 Curr Biol 19(1) :43-9

Bettencourt Dias, M. and Glover (2009). SnapShot: centriole biogenesis Cell 136(1) :188-188.e1

Bettencourt Dias, M. and Goshima (2009). RNAi in Drosophila S2 Cells as a Tool for Studying Cell Cycle Progression Methods Mol Biol 545 :39-62

Cunha-Ferreira I, Bento I, Bettencourt Dias M. (2009). From Zero to Many: Control of Centriole Number in Development and Disease Traffic 10(5) :482-98

Bettencourt Dias, M & Zita Carvalho-Santos (2008). The double life of centrioles: CP110 hits the spotlight Trends in Cell Biology, Research Focus 18(1) :8-11

Rodrigues-Martins A, Riparbelli M, Callaini G, Glover D and Bettencourt Dias, M. (2008). From centriole biogenesis to cellular function: centrioles are essential for cell division at critical developmental stages Cell Cycle 7(1) :11-16

Rodrigues-Martins, A*, Bettencourt Dias, M*, Riparbelli, M*, C Ferreira, I Ferreira, G Callaini & DM Glover (2007). DSAS-6 organizes a tube-like centriole precursor and its absence suggests modularity in centriole assembly Curr Biol 17(17) :1465-1472

Bettencourt Dias, M. and Glover, D.M. (2007). Centrosome Biogenesis and Function Nature Reviews in Molecular and Cellular Biology 8(6) :451-63

Rodrigues-Martins, A., Riparbelli, M., Callaini, G., Glover, D.M. and Bettencourt Dias, M. (2007). Revisiting the role of the mother centriole in centriole duplication.
Recommended by Faculty of 1000 Biology. Science 316(5827) :1046-50 Link

Bettencourt Dias, M., Rodrigues-Martins, A., Carpenter, L., Riparbelli, M., Gatt, M., Lehmann, L., Carmo, N., Balloux, F., Callaini, G., Glover, D.M. (2005). SAK/PLK4 is required for centriole duplication and flagella development. Current Biology 15(24)

Bettencourt Dias, M., Giet, R., Sinka, R., Mazumdar, A., Lock, W.G., Balloux, F., Zafiropoulos, P., Yamaguchi, S., Winter, S., Carthew, R., Cooper, M., Jones, D., Frenz, L. and Glover, D.M. (2004). Genome wide survey of protein kinases required for cell cycle progression.
Selected as a MUST READ paper by Faculty of 1000 Biology Nature 432 :980-7 Link