PXD061395-1

PXD061395 is an original dataset announced via ProteomeXchange.

Title	Role of TbTim20 in mitochondrial protein import in trypanosomes – TbTim20 RNAi data
Description	Protein import across the mitochondrial inner membrane typically depends on two protein translocases of the inner membrane (TIM) complexes and the membrane potential. The protozoan parasite Trypanosoma brucei, however, has a single, divergent TIM complex. Unlike other trypanosomal TIM subunits, TbTim20 is neither essential for normal growth of insect nor bloodstream forms of T. brucei, leaving its role uncertain. Specific mutations in the γ-subunit of the F1FO-ATPase, such as γL262P, permit bloodstream form trypanosomes to grow without mitochondrial DNA (kinetoplast or kDNA). Here we show that RNAi-mediated depletion of TbTim20 inhibits growth of this cell line, but only if it lacks the kDNA. Titration of mitochondrial uncouplers and direct membrane potential measurements reveal that TbTim20 becomes more critical as the membrane potential decreases across all tested lines. Proteomic analysis of the uninduced and induced γL262P TbTim20-RNAi cell line, which lacks kDNA and exhibits the lowest membrane potential, shows depletion of a subset of imported proteins. This subset includes ATPase subunits, explaining why TbTim20-silenced cell lines become more sensitive to uncouplers. Thus, we propose that TbTim20 supports import of a subset of proteins whose import is hypersensitive to a low membrane potential.
HostingRepository	PRIDE
AnnounceDate	2025-10-27
AnnouncementXML	Submission_2025-10-27_02:43:51.011.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Julian Bender
SpeciesList	scientific name: Trypanosoma brucei; NCBI TaxID: NEWT:5691;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2025-03-03 06:04:21	ID requested
⏵ 1	2025-10-27 02:43:51	announced

10.1111/FEBS.70297;

submitter keyword: Trypanosoma brucei,proteomics, mitochondrial genome, RNAi-induced gene depletion, label-free quantification, mitochondrial protein import

Bettina Warscheid
contact affiliation	Chair of Biochemistry II, Theodor Boveri-Institute, Biocenter, University of Würzburg, Würzburg, Germany
contact email	bettina.warscheid@uni-wuerzburg.de
lab head
Julian Bender
contact affiliation	University of Wuerzburg Chair of Biochemistry II Am Hubland 97074 Würzburg
contact email	julian.bender@uni-wuerzburg.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD061395
     1. Label: PRIDE project
     2. Name: Role of TbTim20 in mitochondrial protein import in trypanosomes – TbTim20 RNAi data