Group Leader/s

 

intro

Our research program integrates biochemistry, biophysics, and bottom-up synthetic biology to reconstruct minimal bacterial division machineries from its molecular building blocks in controlled cell-like environments. This effort, which is framed on the quest of building synthetic cells from scratch, will contribute to our understanding of how cells divide and will provide new horizons for biotechnological and biomedical applications.

For more details, please see the personal lab web page at https://mastermcib.wixsite.com/grlabcib

 

BASYC - Bacterial division in synthetic cytomimetic environments
BASYC - Bacterial division in synthetic cytomimetic environments

 

  • We aim at understanding the molecular mechanisms responsible for the biochemical organization of the bacterial division machinery (the divisome). The purpose is to achieve a quantitative description of how FtsZ (the central element of the divisome in most bacteria) and the proteins modulating division ring stability work together as an integrated system of multiple interactions to assemble the functional molecular engine that drives cytokinesis. This biochemical information will contribute to defining more precise conditions to reconstruct different combinations of divisome subsets and to test their functional organization in minimal membrane systems and cell-like containers, as vesicles and droplets produced by microfluidics.
The elements of the bacterial division machinery (the divisome)
The elements of the bacterial division machinery (the divisome): FtsZ - the central protein of the divisome - and the positive / negative regulators of the Z-ring stability in a dividing cell. SHAPE  \* MERGEFORMAT

 

  • Synthetic systems can be devised to mimic elements of the intracellular complexity (as excluded volume effects due to natural crowding, surface interactions, and macromolecular condensation resulting from biologically regulated liquid-liquid phase separation) in media of known and controllable composition.

 

Macromolecular crowding – in vitro, in vivo, and in between.
Macromolecular crowding – in vitro, in vivo, and in between. Diagram illustrating crowding studies of macromolecular reactions - folding and binary hetero-association - in vitro, in vivo and in a synthetic medium of intermediate complexity designed to emulate a particular cellular microenvironment (Rivas and Minton, 2018)

 

  • This integrative approach has allowed us to demonstrate that FtsZ, together with its nucleoid-associated inhibitor SlmA, assembles into dynamic condensates driven by phase separation in crowded cell-like conditions. Our findings suggest that phase-separated biomolecular condensation may also help to organize intracellular space in bacteria.
Macromolecular phase separation in bacterial division.
Macromolecular phase separation in bacterial division. Crowding and phase separation drives the formation of dynamic condensates of FtsZ in the presence of nucleoprotein complexes organized by the FtsZ spatial regulator SlmA. These condensates may constitute a novel element of spatiotemporal organization of essential cell cycle processes in bacteria (Monterroso et al. 2019). SHAPE  \* MERGEFORMAT

 

 

 

Robles-Ramos MÁ, Margolin W, Sobrinos-Sanguino M, Alfonso C, Rivas G, Monterroso B, Zorrilla S.  [2020]. The Nucleoid Occlusion Protein SlmA Binds to Lipid Membranes. mBio. 11(5):e02094-20. doi: 10.1128/mBio.02094-20.

Monterroso B, Zorrilla S, Sobrinos-Sanguino M, Robles-Ramos MA, López-Álvarez M, Margolin W, Keating CD, Rivas G.  [2019]. Bacterial FtsZ protein forms phase-separated condensates with its nucleoid-associated inhibitor SlmA. EMBO Rep. 20(1). pii: e45946. doi: 10.15252/embr.201845946.

Monterroso B, Zorrilla S, Sobrinos-Sanguino M, Robles-Ramos MÁ, Alfonso C, Söderström B, Meiresonne NY, Verheul J, den Blaauwen T, Rivas G.  [2019]. The Bacterial DNA Binding Protein MatP Involved in Linking the Nucleoid Terminal Domain to the Divisome at Midcell Interacts with Lipid Membranes. MBio. 10(3). pii: e00376-19. doi: 10.1128/mBio.00376-19.

Ramirez-Diaz DA, García-Soriano DA, Raso A, Mücksch J, Feingold M, Rivas G, Schwille P.  [2018]. Treadmilling analysis reveals new insights into dynamic FtsZ ring architecture. PLoS Biol. 16(5):e2004845. doi: 10.1371/journal.pbio.2004845.

Sobrinos-Sanguino M, Zorrilla S, Keating CD, Monterroso B, Rivas G.  [2017]. Encapsulation of a compartmentalized cytoplasm mimic within a lipid membrane by microfluidics. Chem Commun (Camb). 53(35):4775-4778. doi: 10.1039/c7cc01289f.

Rivas G, Minton AP.  [2016]. Macromolecular Crowding In Vitro, In Vivo, and In Between. Trends Biochem Sci. 41:970-981.

Martos A, Jiménez M, Rivas G*, Schwille P*  [2012]. Towards a bottom-up reconstitution of bacterial cell division. Trends Cell Biol. 22:634-643

 

Funding

- 2020-2023: BASYC - Bacterial division in synthetic cytomimetic environments. Spanish Goverment. Plan Nacional I+D+i, PID2019-104544GB-I00.

- 2017-2019: BIOROOMS - Analysis, synthesis and reassembly of the biorooms active in bacterial division. Spanish Government Plan Nacional I+D+i, BFU2016-75471-C2-1-P.  

- 2015-2019: MASSTRPLAN - "MASS Spectrometry TRaining network for Protein Lipid adduct Analysis" EU Project 675132 (H2020-MSCA-ITN-2015). Innovative Training Network.

- 2015-2016: ODIVITUBE - Biochemical organization of minimal divisomes in the test tube. Spanish Government, Plan Nacional I+D+i, BFU2014-28941-C03-02.

- 2012-2014: SYNVISION - Synthetic biology of bacterial cell division: reconstruction of minimal divisomes in biomimetic membrane systems. Spanish Government, Plan Nacional I+D+i, BIO2011-28941-C03-03.

- 2011-2014: SYNTHETIC BIOLOGY OF BACTERIAL CELL DIVISION. Human Frontier Science Program RGP0050-2010.

- 2009-2013: DIVINOCELL - Exploiting Gram-negative cell division targets in the test tube to obtain anti-microbial compounds. European Commission, FP7-HEALTH-F3-2009-223431.

 

More info

GR LAB

Centro de Investigaciones Biológicas Margarita Salas - CSIC

Ramiro de Maeztu 9 - E28040 Madrid

Tel. +34 918373112 ext. 4304

E-mail: grivas@cib.csic.es

GRLAB web: http://www.cib.csic.es/en/grupo.grivas & https://mastermcib.wixsite.com/grlabcib

 

HOW TO REACH GRLAB:

http://www.cib.csic.es/es/como_llegar.php?

 

 

 

 

 

  GR lab web: Bioquímica de sistemas de la división bacteriana

  HOW TO REACH GRLAB