Understanding the role of CyclinB1-NuMA interaction in spindle orientation and epithelial morphogenesis

Projektinformationen

NuMA_CyclinB1_EM

ID Finanzhilfevereinbarung: 101028901

DOI

10.3030/101028901

Das Projekt endete am 30 November 2023

EK-Unterschriftsdatum 26 März 2021

Startdatum 1 September 2022

Enddatum 31 August 2024

Finanziert unter

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Gesamtkosten

€ 160 932,48

EU-Beitrag

€ 160 932,48

Koordiniert durch

AGENCIA ESTATAL CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
Spain

Periodic Reporting for period 1 - NuMA_CyclinB1_EM (Understanding the role of CyclinB1-NuMA interaction in spindle orientation and epithelial morphogenesis)

Berichtszeitraum: 2022-09-01 bis 2024-08-31

Zusammenfassung vom Kontext und den Gesamtzielen des Projekts

Cell division is controlled not only in time, but also in orientation. Our aim is to advance knowledge on how cell cycle regulates spindle positioning, crucial to orientate cell division. Specifically, this research aims to elucidate the role of CyclinB1 in the formation and orientation of the mitotic spindle and its contribution to epithelial morphogenesis through its interaction with NuMA. Therefore, our mains objectives are finding how, when, where and why CyclinB1 and NuMA interact.

I propose a multidisciplinary project to understand better the molecular mechanisms associated with cell division, spindle regulation, and its implication in epithelial morphogenesis. The cell cycle is intimately involved in all biological organisms' growth, development, reproduction, and aging; therefore, gaining insight into cell cycle controls will benefit the large community of biologists and medical sciences. Furthermore, establishing the correct orientation of the mitotic spindle is an essential step in epithelial cell division to ensure that epithelia tubes from correctly during organ development and regeneration. Disrupting of any of the spindle factors, could lead to aneuploidy, epithelial to mesenchymal transition and tissue disorganization. Moreover, a better understanding of mammalian cell cycle controls has significant potential to change medical treatments for cancer cells that have escaped the normal controls on growth and division in multicellular animals. Providing novel insights into cell cycle regulation and its implication in epithelial morphogenesis could potentially lead to therapies and treatments that could dramatically improve the economy's well-being, health, and creative output. Potential public sector health benefits would be realized over a more extended period.

Arbeit, die ab Beginn des Projekts bis zum Ende des durch den Bericht erfassten Berichtszeitraums geleistet wurde, und die wichtigsten bis dahin erzielten Ergebnisse

The Mass Spectrometry data obtained from endogenous tagged NuMA CRISPR/Cas9 cells shows that CyclinB1/CDK1 are two of the main NuMA interactors in mitosis. Spatial and temporal localization of both proteins were analyzed in CyclinB1-NuMA CRISPR edited cells in vivo, in situ (PLA) and in vitro (immunoprecipitation). The results show that CyclinB1-NuMA interaction takes first place in the nucleus in prophase before the nuclear envelope breaks. In prometaphase and metaphase, both proteins colocalizes and interacts at the spindle poles showing a dynamic behavior by FRAP. By mutagenesis and immunoprecipitation approaches, two motifs in NuMA tail were identified as essential for CyclinB1 interaction. Finally, CyclinB1 fails to localize to the mitotic spindle in NuMA depleted cells. This result suggests that NuMA could be the spindle pole receptor of CyclinB1. Therefore, Cyclin B1/CDK1 could be an integral component besides an essential regulator of the mitotic spindle through its interaction with NuMA at the spindle poles.

The results of this project have been disseminated through regular laboratory meetings and seminars. I presented this work at the international EMBO workshop cell polarity and membrane dynamics, organized by EMBO (21/05/2023-26/05/2023, Girona, Spain). At that workshop, I was invited to give a short communication, titled: “Looking for the role of CyclinB1 in spindle orientation”. I also presented a poster. I have participated in the European Research Night organized by CSIC (09/30/2022, Madrid, Spain) with the activity “Nobel and Non-Nobel Women”. During the activity, the aim was to transmit and give visibility to the general public about the role of women scientists who had received a Nobel Prize throughout history, and about other researchers whose authorship had remained in the shadows. Moreover, I have been invited to give a talk at Salesianos High School (April 2024, Madrid, Spain). As a MSCA fellow, I won the third prize in the Huygens Image contest 2023 (https://svi.nl/ImageContestWinner) with a picture of a human cell in metaphase. The head of the communications and public engagement team at the CBMSO spread the information on social media (Twitter, Facebook and Instagram).

Fortschritte, die über den aktuellen Stand der Technik hinausgehen und voraussichtliche potenzielle Auswirkungen (einschließlich der bis dato erzielten sozioökonomischen Auswirkungen und weiter gefassten gesellschaftlichen Auswirkungen des Projekts)

This project aims to uncover the essential role that CyclinB1 plays in the regulation of the mitotic spindle through its interaction with NuMA and the implication of this interaction in epithelial morphogenesis. For just over a year, we have discovered that CyclinB1, and its kinase CDK1, are two of the main interactors of NuMA in mitosis. CyclinB1 and NuMA interacts in prophase in the nucleus, and prometaphase /metaphase at the spindle poles showing a dynamic behavior. Moreover, two motifs in NuMA have been identified as essential for CyclinB1 interaction. Interestingly, CyclinB1 fails to localize to the mitotic spindle in NuMA depleted cells. This result suggests that NuMA could be the spindle pole receptor of CyclinB1. Therefore, Cyclin B1/CDK1 could be an integral component besides an essential regulator of the mitotic spindle through its interaction with NuMA at the spindle poles.

Our research would impact (1) academic researchers studying cell cycle, epithelial morphogenesis, and cancer, (2) industrial organizations devising therapies and (3) public benefits by improving the quality of life.

Human cell in metaphase. During this stage of cell division, condensed chromosomes line up at the me