PXD043570

PXD043570 is an original dataset announced via ProteomeXchange.

Title	Quantitative proteomic analysis based on the isobaric tags for relative and absolute quantitation (iTRAQ) labeling in WT and NudCL2 KO-1 HEK-293 cells
Description	Cytokinesis is a key step in the later stage of cell division to partition cellular contents into daughter cells, which undergoes precise regulation in temporal and spatial. However, the underlying mechanisms are still unclear. Here, we show that Hsp90 cochaperone, NudCL2 (NudC-like protein 2), is required for cytokinesis in mammalian cells. NudCL2 localizes to midbody during cytokinesis. Knockout (KO) of NudCL2 using CRISPR/Cas9-based genome editing causes cytokinesis failure and leads to the accumulation of multinucleated cells. To investigate the potential regulator involved in NudCL2-mediated cytokinesis regulation, we performed isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic analysis in WT and NudCL2 KO cells and found hundreds of differentially expressed proteins (KO/WT fold change > 1.2 or < 0.83, p < 0.05) in NudCL2 KO cells. We found that the protein level of RCC2 (a midbody-associated protein) is obviously decrease in NudCL2 KO cells. Further studies display that knockout of NudCL2 induces an increase in RCC2 protein instability and degradation. Our data show that depletion of RCC2 leads to the similar defects as NudCL2 KO. Ectopic expression of RCC2 effectively rescues the cytokinesis failure caused by the loss of NudCL2. Interestingly, our data reveals that Hsp90 localizes to midbody in cytokinesis, and interacts with NudCL2 and RCC2. Inhibition of Hsp90 ATPase activity also causes the RCC2 instability and cytokinesis failure as NudCL2 KO. Moreover, the abnormal phenotypes induced by NudCL2 KO are able to rescue by the ectopic expression of Hsp90, but not vice versa. Taken together, our data suggest that NudCL2 is required to cytokinesis by stabilizing RCC2 at midbody through the Hsp90 pathway, providing a hitherto unrecognized mechanism crucial for cytokinesis regulation.
HostingRepository	PRIDE
AnnounceDate	2024-03-21
AnnouncementXML	Submission_2024-03-21_01:30:06.988.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Xiaoyang Xu
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2023-07-05 21:42:28	ID requested
⏵ 1	2024-03-21 01:30:07	announced

Dataset with its publication pending

submitter keyword: HEK-293, NudCL2, iTRAQ

Tianhua Zhou
contact affiliation	Department of Cell Biology, and Institute of Gastroenterology of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China
contact email	tzhou@zju.edu.cn
lab head
Xiaoyang Xu
contact affiliation	Zhejiang University School of Medicine
contact email	1273393591@qq.com
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/2024/03/PXD043570

PRIDE project URI

• PRIDE
  1. PXD043570
     1. Label: PRIDE project
     2. Name: Quantitative proteomic analysis based on the isobaric tags for relative and absolute quantitation (iTRAQ) labeling in WT and NudCL2 KO-1 HEK-293 cells