Our laboratory is interested in three main areas: 1. How centrosomes and cilia are built and maintained; 2. How centrioles and cilia have evolved; and 3. How centrosomes and cilia are deregulated in disease.

1.How centrosomes and cilia are built and maintained

Centrosome Biogenesis in Space and Time– The formation of new centrioles is highly regulated in animal cycling cells: once every cell cycle, one new “daughter” centriole grows close to the already existing one, the “mother”, ensuring maintenance of centriole number. Centriole biogenesis raises many interesting questions also relevant for other structures in the cell: how is time, place and number regulated. We identified PLK4, a critical trigger for centriole formation (Bettencourt-Dias et al, 2004; 2005). We further showed that centriole biogenesis does not always depend on a template centriole but can originate de novo, upon PLK4 overexpression (Rodrigues-Martins et al, 2007; Lopes et al, 2015; Nabais et al, 2021). De novo formation relies on the same components as canonical biogenesis (Rodrigues-Martins et al, 2007; Pereira et al, 2021). We discovered mechanisms regulating centriole biogenesis in space and time, through positive and negative feedback loops controlling the activity of PLK4 and other components. The number of centrioles formed relies on control of PLK4 levels, through negative feedback loops, protein degradation and cell cycle control of PLK4 by CDK1 (Cunha-Ferreira et al, 2009, 2013; Zitouni et al, 2016; Gouveia et al, 2018).

Centrosome and Cilia Maintenance– Centriole inheritance and function depend on their ability to persist through many cell cycles and in long-lived cells. Centrioles are known to be remarkably stable: in contrast to cytoplasmic MTs, they are resistant to drug and cold-induced MT depolymerization. We uncovered a centrosome-maintenance mechanism that is shut down in the female germline. Loss of the pericentriolar material (PCM), triggered by down regulation of the essential PCM recruiter Polo-kinase (PLK1 in humans), leads to centriole elimination. Artificial tethering of Polo to centrosomes, led to PCM maintenance and prevented centriole loss, indicating PLK1 is sufficient to ensure centriole maintenance. Regulation of this program is key to sexual reproduction, as artificial maintenance of centrioles led to abnormal meiotic division, followed by aborted zygotic development (Pimenta-Marques et al, 2016). Our recent data suggests that the PCM plays a critical role in maintaining high levels of essential centriole proteins, such as ANA1 (Pimenta-Marques and Perestrelo et al, 2024). This work showed that centrioles are not intrinsically stable, and that the PCM and centriole components are critical for their maintenance. Moreover, this work suggested that centriole components are more dynamic than previously thought. We are investigating how the PCM and centriole components provide stability to the centrioles, whether this is differently regulated in different cell types and the possible consequences of its mysregulation for the organism. Moreover, we are also investigating how basal body and cilia maintenance is regulated in Drosophila sensory neurons. We uncovered several critical properties of centrioles by investigating centrioles and cilia in fruit flies and human non-transformed and cancer cells. We found that centriole structure varies and adapts for different functions, explaining why mutations in conserved centriole components lead to tissue-specific diseases, particularly ciliopathies (Jana et al, 2018; Chen et al, 2015). Moreover, we also uncovered that cilia functional maintenance depends on key regulatory molecules such as IFT88 (Werner and Ramos et al, 2024). Our results offer an explanation to how mutations in conserved centriole components lead to tissue-specific disease.

2.How centrioles and cilia have evolved

Driven by our deep interest in evolution, our group ventured into the emerging field of evolutionary cell biology. Combining comparative genomics and cell biology, we identified the ancestral centriole in LECA (Last Eukaryotic Common Ancestor) and traced their step-wise evolution, incorporating new regulatory elements (Carvalho-Santos et al, 2010; Carvalho-Santos et al, 2011; Ito et al, 2019; Ito et al, 2019). We continue to explore the evolutionary history and diversity of the basal body cytoskeleton, contributing to a broader understanding of the evolution of cellular complexity.

3.How centrosomes and cilia are deregulated in disease

Centrosome and cilia alterations, in particular the presence of multiple centrioles, can lead to abnormalities in chromosome segregation, asymmetric cell divisions, invasiveness and ability to induce cancer in mice. Multiple questions remain, in particular how those changes originate, how they contribute to cancer and how can they be taken advantage off for translational purposes. We use different experimental systems to tackle those questions. We established centriole amplification and size deregulation as recurrent features of cancer cells, that can occur early in tumor progression, are dependent on the loss of p53 and are associated with bad prognosis. Moreover, we identified novel causes and consequences of those abnormalities. Abnormalities in centrosome number and structure were reported in many clinical samples. However, our understanding of the prevalence, origins and consequences of such defects was limited.  We showed that centrosome alterations can appear in pre-malignant condition and expand in neoplasia, being associated with worse outcomes (Lopes et al, 2018; Marteil et al, 2018), suggesting they have an important role in tumor progression. Moreover, we identified a pan-cancer association of a centrosome amplification gene expression signature with genomic alterations and clinical outcome (Almeida et al, 2019). We suggest how cells with centrosome amplification may be kept within the tumour population, resisting negative selection (Louro et al, 2021) and how coping mechanisms might be critical for survival of tumors with low epithelial characteristics, in particular in leukemias (https://www.biorxiv.org/content/10.1101/2023.03.13.532472v1). Using cancer cell lines, including the NCI-60 panel of human cancer cell lines derived from 9 distinct tissues, we continue to address the highlighted questions combined with bioinformatics and modelling of centriole number along tumor progression.

Biogenesis and function

1.PROJETO PTDC/BIA-CEL/4202/2021

This project investigates the regulation of centriole de novo biogenesis and its role in tumorigenesis, for which there is less information available.

2. MOORE FOUNDATION- EVOLUTION OF CENTRIOLES, CILIA AND MEIOSIS. Our lab continues to explore the evolutionary history and diversity of the basal body cytoskeleton, contributing to a broader understanding of the evolution of cellular complexity. We seek to explore the profound evolutionary transitions from prokaryotic to eukaryotic cells, focusing specifically on the role of basal bodies and microtubule organizing centers (MTOCs) in fundamental eukaryotic processes such as mitosis and meiosis. Basal bodies/Centrioles are highly conserved across eukaryotes. However, they show variability in their function and structure across different taxa and life stages, including their role in both chromosomal segregation and the organization of flagella. We propose basal bodies’ original functions were likely centered around flagellar dynamics, with their roles in division developing secondarily, potentially multiple times across different evolutionary paths. This project is funded by the Moore Foundation and harnesses a diverse team to explore the complex roles of basal bodies in eukaryotic cells, focusing on their evolutionary significance and variability across different taxa and life stages. The team comprises experts from several labs: P. Keeling Lab (UBC, Canada) known for its expertise in target organisms and evolutionary analyses, H. Goodson Lab (UND, USA), offering knowledge in the evolution of cytoskeletal systems, and J. Wideman Lab (UA, USA), specializing in spatial proteomics. Outcomes of this collaborative effort include developing a detailed understanding of the structural and functional evolution of basal bodies and their roles in crucial cellular processes like meiosis and mitosis. The integration of these findings is expected to offer new perspectives on the origin and evolution of key cellular functions, contributing significantly to the field of evolutionary cell biology.

A. Pimenta-Marques, T. Perestrelo, […] M. Bettencourt-Dias** (2024). Ana1/CEP295 is an essential player in the centrosome maintenance program regulated by Polo kinase and the PCM. EMBO Reports 25(1): 102-127.
Contribution:Identified centriole components important for centriole maintenance.

S. Werner*, P. Okenve-Ramos, […] M.C. Göpfert**, S.C. Jana**, M. Bettencourt-Dias** (2024). IFT88 maintains sensory function by localizing signaling proteins along Drosophila cilia. Life Science Alliance 7(5): e202302289. *co-first authors; *co-senior authors.
Contribution: Showed that sensory ciliary function underlying hearing in the adult fly requires an active maintenance program involving DmIFT88 and its signaling transmembrane cargoes, DmGucy2d and Inactive.

S. Gomes Pereira*, […] J.D. Becker**, M. Bettencourt-Dias** (2021). 3D architecture and molecular foundations of de novo centriole assembly via bicentrioles. Current Biology 31(19): 4340-4353.e7. *co-first authors; *co-senior authors.
Contribution: Used mosses to mechanistically study the bicentriole pathway for de novo centriole assembly, showing shared players between de novo and canonical centriole biogenesis.

C. Nabais*, D. Pessoa, J. de-Carvalho, T. van Zanten, P. Duarte, S. Mayor, J. Carneiro, I.A. Telley**, M. Bettencourt-Dias** (2021). Plk4 triggers autonomous de novo centriole biogenesis and maturation. Journal of Cell Biology 220(5): e202008090. *co-first authors; *co-senior authors.
Contribution: Showed that gamma-tubulin is essential for de novo centriole biogenesis in a Drosophila egg extract system.

D. Ito*, […] M. Bettencourt-Dias** (2019). Pericentrin-mediated SAS-6 recruitment promotes centriole assembly. eLife8: e41418. *co-first authors; *co-senior authors.
Contribution: Discovered the importance of PCM in recruiting centriole components using yeast as a model.

S.M. Gouveia*, S. Zitouni, […] M. Bettencourt-Dias** (2019). PLK4 is a microtubule-associated protein that self-assembles, promoting de novo MTOC formation. Journal of Cell Science 132(4): jcs219501. *co-first authors; *co-senior authors. Contribution: Demonstrated that PLK4 forms condensates recruiting other centriole and PCM components.

S.W. Jana*, […] M. Bettencourt-Dias** (2018). Differential regulation of transition zone and centriole proteins contributes to ciliary base diversity. Nature Cell Biology 20(8): 928-941. *co-first authors; *co-senior authors.
Contribution: Created a structural and biochemical atlas of the ciliary base, uncovering mechanisms for tissue-specific diseases caused by centriole mutations.

G. Marteil*, […] M. Bettencourt-Dias** (2018). Over-elongation of centrioles in cancer promotes centriole amplification and chromosome missegregation. Nature Communications 9(1): 1-17. *co-first authors; *co-senior authors.
Contribution: Established that centriole amplification and size deregulation are features of cancer associated with poor prognosis.

A. Pimenta-Marques**, I. Bento**, […] M. Bettencourt-Dias** (2016). Mechanism for the elimination of the female gamete centrosome in Drosophila melanogaster. Science 353(6294): aaf4866. co-senior authors. Contribution: Found that centrioles can disassemble, revealing a PLK1-dependent program ensuring centriole maintenance.

C.A. Lopes*, S.C. Jana*, I. Cunha-Ferreira, […] M. Bettencourt-Dias** (2015). PLK4 trans-autoactivation controls centriole biogenesis in space. Developmental Cell 35(2): 222-235. *co-first authors; *co-senior authors.
Contribution:Demonstrated that centrioles promote their own reproduction via PLK4 autoactivation.

A. Rodrigues-Martins, […] M. Bettencourt-Dias** (2007). Revisiting the role of the mother centriole in centriole biogenesis. Science 316(5827): 1046-1050.
Contribution: Showed that centrioles can form de novo and self-assemble without a template.

2024                     Membership of the Portuguese Academy of Sciences

2024                     Merit Medal by the Portuguese Foundation for Science & Technology

2022                     Gold Medal of Merit by Oeiras City Council

2017-2023              ERC Consolidator Grant (PI)- Centriole Birth and Death

2015                     EMBO membership

2012                     Keith R. Porter Fellowship (Philadelphia Foundation and ASCB)

2011-2016            ERC Starting Grant (PI)- Control of Centriole Structure&Number

2009                     EMBO Young Investigator Programme

2009                     Schlumberger Fondation Prize

2007-2012            EMBO Installation Grant (PI) – Centriole Biogenesis

2007 & 2012         Pfizer Award for Basic research

2007                     Eppendorf European Young Investigator