Functional reconstitution of molecular machines in microdroplets allows to discern the effect of physiological factors such as macromolecular crowding,microenvironments and membranes in their operation The encapsulation of different combinations of elements of the E coli division machinery using microfluidics technology evidenced their structural and functional organization in droplets where the boundary mimics the lipid composition of the intracellular membrane We could determine the possible role of the membrane in the assembly and localization of biomolecular condensates formed by FtsZ (GTPase key in cell division), SlmA (effector of the nucleoid occlusion system negatively regulating Z ring assembly) and the DNA sequences specifically recognized by the latter.The location of the biomolecular condensates at the membrane seems to be modulated by their composition Thus, for these heterotypic condensates it is possibly related with the membrane binding of SlmA we identified being maintained in the presence of a positive regulator of the Z ring assembly counteracting the effect of SlmA and basically lost in homotypic FtsZ condensates 4 Additionally, we could characterize the differential distribution of biomolecular condensates and FtsZ polymers resulting from their nucleotide dependent switch, both in homogeneous crowding and in intracellular compartmentation models using binary phase. Our results evidence the advantages of using this kind of technological approximations to identify new players within the biomolecular interaction networks and better characterize, in a quantifiable and reproducible manner,their effects.

info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2022-136951NB-I00/ES/RECONSTITUCION DE LA DIVISION CELULAR EN SISTEMAS MINIMOS CITOMIMETICOS/

MICINN(PID2019-104544-GB-I00/10 13039 501100011033 PID2022-136951-NB-I00 funded by MCIN/AEI/ 10 13039 501100011033 and ERDF “A way of making Europe”

Peer reviewed

1 p.-5 fig.