PXD069854

PXD069854 is an original dataset announced via ProteomeXchange.

Title	A ZNF-nanobody fusion reveals SUMOylation-dependent changes in protein localization
Description	SUMOylation is a post-translational modification regulating protein localization, stability, and activity, with effects varying depending on the identity of the conjugated SUMO protein (SUMO1 or SUMO2/3) and type of SUMOylation (mono-, multi-, or poly-). We previously developed a small 32 amino acids SUMOylation tag derived from the ZNF451 E3 ligase (hereafter “ZNF”) that biases substrates toward SUMO2/3 conjugation and modulates the transcriptional activity of p53. To determine how enhanced SUMOylation affects protein localization, we fused ZNF to the GFP-binding nanobody vhhGFP4 (VHH), creating VHH-ZNF to drive SUMOylation of GFP-tagged substrates in trans. In vitro, VHH-ZNF increased SUMO2/3 modification of p53-GFP, preferentially generating polySUMO2 chains at the canonical K386 site; using a chain-deficient SUMO2(K0) shifted the pattern toward multi-SUMOylation. VHH-ZNF did not modify HA-p53 alone but SUMOylated it in the presence of p53-GFP, consistent with proximity-based modification within p53 tetramers. In HEK293 cells co-expressing p53-GFP and VHH-ZNF, immunoblotting and proteomics confirmed increased SUMO2/3 conjugation at K386 that was abrogated by the E1 inhibitor ML792. Fluorescence microscopy analyses revealed that SUMOylated p53 transitions from a diffuse nuclear distribution to SUMO-positive nuclear foci that partially overlap with promyelocytic leukemia (PML) nuclear bodies. Overall, we developed a new method to increase the SUMOylation of GFP-tagged proteins and to visualize SUMOylation-dependent relocalization in cells.
HostingRepository	PRIDE
AnnounceDate	2026-03-24
AnnouncementXML	Submission_2026-03-24_06:19:57.944.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Chongyang Li
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	phosphorylated residue; sumoylated lysine; monohydroxylated residue; deamidated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2025-10-23 17:08:55	ID requested
⏵ 1	2026-03-24 06:19:58	announced

Dataset with its publication pending

submitter keyword: None

Pierre Thibault
contact affiliation	1. Institut de Recherche en Immunologie et Cancérologie (IRIC), Université de Montréal, Montréal, Québec, Canada. 2. Département de Biochimie et Biologie Moléculaire, Université de Montréal, Montréal, Québec, Canada. 3. Département de Chimie, Université de Montréal, Montréal, Québec, Canada
contact email	pierre.thibault@umontreal.ca
lab head
Chongyang Li
contact affiliation	Institute for Research In Immunology and Cancer, University of Montreal
contact email	chongyang.li@umontreal.ca
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD069854

PRIDE project URI

• PRIDE
  1. PXD069854
     1. Label: PRIDE project
     2. Name: A ZNF-nanobody fusion reveals SUMOylation-dependent changes in protein localization