PXD068118

PXD068118 is an original dataset announced via ProteomeXchange.

Title	Tracking Arabidopsis PCNA1 and PCNA2 with a Human PCNA Nanobody-Based Chromobody
Description	Background Proliferating Cell Nuclear Antigen (PCNA) is a crucial protein in eukaryotes that recruits nuclear factors during DNA replication and repair, but also cytoplasmic proteins involved in apoptosis and cell signaling. In the model plant Arabidopsis thaliana, two highly similar PCNA isoforms, AtPCNA1 and AtPCNA2, have been identifi ed. However their functions remain unexplored, probably because loss-of-function mutants, as observed in other model systems, are lethal. The use of nanobodies -single-domain antibody fragments- for targeted protein degradation and mislocalization represents a promising complementary alternative for studying gene function. However, these approaches are still poorly used in plant research. Results To explore the applicability of this strategy in plants, we designed and expressed a modifi ed human PCNA chromobody (PCNA Cb) consisting of a commercial anti-human PCNA nanobody-fl uorescent protein fusion, in Arabidopsis to test its capacity to track the dynamics of PCNA isoforms. We fi rst demonstrated that AtPCNA1 and AtPCNA2 assemble in a heteromeric complex and exhibit a highly overlapping localization during the cell cycle. Co-immunoprecipitation assays and microscopic analyses confi rmed that PCNA Cb binds to AtPCNA1 and AtPCNA2 in vivo and fully colocalizes with them throughout the cell cycle in living cells. Conclusions Our results demonstrate that a human-derived PCNA chromobody can be used to monitor endogenous Arabidopsis PCNA proteins. While further optimization is required to enhance binding effi ciency, this tool offers a promising platform for future studies aimed at elucidating PCNA function in plants.
HostingRepository	PRIDE
AnnounceDate	2025-09-11
AnnouncementXML	Submission_2025-09-11_01:55:27.001.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD068118
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Johana Chicher
SpeciesList	scientific name: Arabidopsis thaliana (Mouse-ear cress); NCBI TaxID: 3702;
ModificationList	phosphorylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	timsTOF Pro 2

Revision	Datetime	Status	ChangeLog Entry
0	2025-09-05 03:15:25	ID requested
⏵ 1	2025-09-11 01:55:27	announced

10.6019/PXD068118;

submitter keyword: Arabidopsis,PCNA, live-imaging, chromobody, nanobody, DNA replication

Mathieu Ingouff
contact affiliation	DIADE, IRD, CIRAD, Université de Montpellier, Montpellier, France
contact email	mathieu.Ingouff@ird.fr
lab head
Johana Chicher
contact affiliation	Université de Strasbourg, Plateforme Protéomique Strasbourg-Esplanade, Centre National de la Recherche Scientifique
contact email	j.chicher@ibmc-cnrs.unistra.fr
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/2025/09/PXD068118

PRIDE project URI

• PRIDE
  1. PXD068118
     1. Label: PRIDE project
     2. Name: Tracking Arabidopsis PCNA1 and PCNA2 with a Human PCNA Nanobody-Based Chromobody