Proximity labelling of mitochondria and endosomes upon mtDNA replication stress.

ABSTRACT: This project aims to identify novel regulators of mtDNA turnover, for what we used TurboID. For specific detection of the mitochondria-endosome proteome, we fused the N-terminal part of Turbo ID to RAB5C (TurboID aa1-aa72; RAB5C-SplitTurboNt-V5) and, the C-terminal part to SAMM50 (TurboID aa73–aa246; SAMM50-SplitTurboCt-HA). Proximity biotinylation was performed in transduced HEK293 cells expressing Split TurboID constructs. Cells expressing only SAMM50-SplitC-HA were used as a negative control. For protein isolation, HEK293 cells were incubated for 4 h with 0,25 mM biotin. For inducing mtDNA damage, cells were previously transfected with Twinkle K319E-Cherry 24 h before the biotin incubation.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: David Pla Martin  

LAB HEAD: David Pla Martín

PROVIDER: PXD049081 | Pride | 2025-04-07

REPOSITORIES: Pride