P53 Sumoylation
P53 Sumoylation - Related Videos
06:00
SUMOylation Assay: An In Vitro Technique to Detect the SUMOylation Status of Substrate Proteins by Immunoblotting
0 Views •
2025
This video demonstrates the in vitro method for SUMOylation of substrate proteins using a sequential enzyme cascade. Further, the SUMOylated status of the protein is identified using the electrophoresis and immunoblotting technique.
09:40
In Vivo Detection and Analysis of Rb Protein SUMOylation in Human Cells
0 Views •
Cited by 2 •
2017
Small ubiquitin-related modifier (SUMO) family proteins are conjugated to the lysine residues of target proteins to regulate various cellular processes. This paper describes a protocol for the detection of retinoblastoma (Rb) protein SUMOylation under endogenous and exogenous conditions in human cells.
04:56
Detection of Aggregation-Prone Behavior in Mutant P53 V157F Breast Cancer Cells Using Multipoint Thioflavin T Fluorescence
0 Views •
2025
Hs578T breast cancer cells harboring the p53 V157F mutation exhibit significantly higher Thioflavin T fluorescence compared to MCF7 cells, indicating enhanced protein aggregation. Multipoint fluorescence measurements improve detection accuracy and reliability in identifying β-sheet-rich aggregates, underscoring the importance of aggregation-prone p53 mutations in cancer research and the development of therapeutic strategies.
10:55
Purification of Ubiquitinated p53 Proteins from Mammalian Cells
0 Views •
Cited by 1 •
2022
The protocol describes a step-by-step method to purify ubiquitinated proteins from mammalian cells using the p53 tumor suppressor protein as an example. Ubiquitinated p53 proteins were purified from cells under stringent nondenaturing and denaturing conditions.
09:45
In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
0 Views •
Cited by 11 •
2018
Unlike ubiquitin ligases, few E3 SUMO ligases have been identified. This modified in vitro SUMOylation protocol is able to identify novel SUMO E3 ligases by an in vitro reconstitution assay.
FAQs
The pathway proceeds through SUMO activation, conjugation, and transfer to p53 by a SUMO E3 ligase. Each stage contributes to whether modification occurs and which p53 lysine residues receive SUMO. This layered control allows cellular conditions, including stress, to influence the extent and consequences of p53 regulation rather than treating sumoylation as a single-step event.
SUMO proteases provide reversibility by removing SUMO from p53 after attachment. Their activity can therefore change the duration or level of the modified state, allowing p53 regulation to remain dynamic during cellular stress. Considering both conjugation and deconjugation is essential when interpreting how sumoylation affects p53-dependent responses.
SUMO attachment can influence several functionally distinct properties of p53, including its stability, subcellular localization, protein interactions, and transcriptional activity. These changes may alter how p53 coordinates cell-cycle arrest, DNA-damage responses, senescence, or apoptosis. The outcome therefore depends on how modification changes p53 regulation and its interactions within the cell.
The major outcomes to examine are changes in p53-dependent transcription and the resulting cellular responses. These include cell-cycle arrest, responses to DNA damage, senescence, and apoptosis. Tracking this range is important because sumoylation may affect p53 at multiple regulatory levels, linking a molecular modification to broader decisions about whether stressed cells stop dividing, enter senescence, or undergo programmed cell death.
A useful study framework includes the SUMO activation and conjugation steps, the E3 ligase-mediated transfer to p53, and SUMO proteases that reverse the modification. It should also evaluate consequences for p53 stability, localization, interactions, and transcriptional activity. Examining these components together helps connect pathway regulation with the cellular response to stress.
P53 sumoylation is relevant because it can modify the activity of a tumor suppressor that coordinates several protective responses to cellular stress. Changes in this regulatory pathway may influence cell-cycle control, DNA-damage responses, senescence, and apoptosis. Studying those links helps researchers investigate mechanisms governing p53-dependent tumor suppression and their significance in cancer biology.