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Whole cell proteome of HeLa Flp in T-RexTM RNF4 WT treated with either CCW16, 1a or 2c

ABSTRACT: The SUMO-targeted E3 ubiquitin ligase RNF4 plays an important role in safeguarding genome and proteome integrity. RNF4 recognizes polySUMO modified proteins and induces their proteolytic or non-proteolytic ubiquitylation. Based on the key functions of RNF4 maintaining proteome and genome stability, control of DNA damage response and DNA-protein crosslink repair, we explored its role in cancer cell survival and as a potential drug target. We found that RNF4 is overexpressed in acute myeloid leukemia and expression levels correlate with poor survival. To develop PROTACs for targeted degradation of RNF4, we aimed to exploit the recently developed cysteine-reactive RNF4 ligand CCW16, and synthesized a set of CCW16-derived degrader molecules using established VHL- and CRBN E3 ligands. We showed that treatment with two CCW16-derived PROTACs leads to overexpression of HMOX1, a ferroptosis marker, and HSP70 protein family members. However, we could not detect a significant reduction of RNF4 levels. Interestingly, also the RNF4 binder CCW16 itself induced the expression of HMOX1. Altogether our data indicates that treatment with CCW16 leads to a ferroptotic cell death pathway.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hela Cell

SUBMITTER: Stefan Müller

LAB HEAD: Prof. Dr. Stefan Müller

PROVIDER: PXD056951 | Pride | 2025-10-13

REPOSITORIES: pride

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			Action	DRS
	20230707_LU_LC2_GT_GG_E114_400ng_F.msf	Msf
	20230707_LU_LC2_GT_GG_F01.raw	Raw
	20230707_LU_LC2_GT_GG_F02.raw	Raw
	20230707_LU_LC2_GT_GG_F03.raw	Raw
	20230707_LU_LC2_GT_GG_F04.raw	Raw

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