PXD000252

PXD000252 is an original dataset announced via ProteomeXchange.

Title	Granzyme M targets topoisomerase II alpha to trigger cell cycle arrest and caspase-dependent apoptosis in cancer cells
Description	Cytotoxic lymphocyte protease granzyme M (GrM) is a potent inducer of tumor cell death in vitro and in vivo. The apoptotic phenotype and mechanism by which it induces cell death however, remain poorly understood and controversial. Here, we show that GrM-induced cell death was largely caspase-dependent with various hallmarks of classical apoptosis, coinciding with caspase-independent G2/M cell cycle arrest. Using positional proteomics in HeLa cells, we identified the nuclear enzyme topoisomerase II alpha (topoII alpha) as a physiological substrate of GrM. Cleavage of topoII alpha by GrM at Leu1280 dissected topoII aplha functional domains from the nuclear localization signals, leading to nuclear exit of topoII alpha catalytic activity into the cytoplasm, and rendering it nonfunctional. Similar to the apoptotic phenotype of GrM, topoII aplha depletion in tumor cells led to cell cycle arrest in G2/M, mitochondrial perturbations, caspase activation, and apoptosis. We conclude that cytotoxic lymphocyte GrM targets topoII aplha to trigger cell cycle arrest and caspase-dependent apoptosis. Raw data files were processed with Mascot distiller 2.3. The mgf files were searched with Mascot daemon 2.3. The quantification was also done by Mascot Distiller. All data was stored in ms_lims. Fixed modifications: methionine oxidation, trideutero-acetylation of lysine, carbamidomethylation of cysteine. Variable modifications: acetylation of peptide N-terminus, trideutero-acetylation of peptide N-terminus, pyroglutamate formation of N-terminal glutamine Enzyme: semi-Arg-C/P with one missed cleavage allowed. Precursor mass tolerance: 10 ppm. Peptide fragment mass tolerance: 0.5 Da Quantitation method: triple SILAC arginine +6 Da and +10 Da
HostingRepository	PRIDE
AnnounceDate	2014-07-28
AnnouncementXML	Submission_2014-07-28_00:36:56.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD000252
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Pieter-Jan De Bock
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	L-methionine sulfoxide: 15.994915; 6x(13)C: 4x(15)N labeled L-arginine; 2-pyrrolidone-5-carboxylic acid (Gln): -17.026549; 6x(13)C labeled residue: 6.020129; iodoacetamide derivatized residue: 57.021464; N-acetylated residue: 42.010565; acetate labeling reagent (N-term) (heavy form: +3amu)
Instrument	LTQ Orbitrap; instrument model: LTQ-Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2013-05-13 03:01:14	ID requested
1	2013-11-27 08:45:16	announced
1	2013-11-28 01:41:45	announced
⏵ 2	2014-07-28 00:36:56	announced	Updated project metadata.

Dataset with its publication pending

submitter keyword: Human, granzyme M, LC-MSMS

Pieter-Jan De Bock
contact affiliation	UGent VIB department of medical protein research
contact email	pieterj.debock@ugent.be
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD000252
     1. Label: PRIDE project
     2. Name: Granzyme M targets topoisomerase II alpha to trigger cell cycle arrest and caspase-dependent apoptosis in cancer cells