A protein folding molecular imaging biosensor monitors the effects of drugs that restore mutant p53 structure and its downstream function in glioblastoma cells

Oncotarget
Ramasamy PaulmuruganTarik F Massoud

Abstract

Misfolding mutations in the DNA-binding domain of p53 alter its conformation, affecting the efficiency with which it binds to chromatin to regulate target gene expression and cell cycle checkpoint functions in many cancers, including glioblastoma. Small molecule drugs that recover misfolded p53 structure and function may improve chemotherapy by activating p53-mediated senescence. We constructed and optimized a split Renilla luciferase (RLUC) complementation molecular biosensor (NRLUC-p53-CRLUC) to determine small molecule-meditated folding changes in p53 protein. After initial evaluation of the biosensor in three different cells lines, we engineered endogenously p53P98L mutant (i.e. not affecting the DNA-binding domain) Ln229 glioblastoma cells, to express the biosensor containing one of four different p53 proteins: p53wt, p53Y220C, p53G245S and p53R282W. We evaluated the consequent phenotypic changes in these four variant cells as well as the parental cells after exposure to PhiKan083 and SCH529074, drugs previously reported to activate mutant p53 folding. Specifically, we measured induced RLUC complementation and consequent therapeutic response. Upon stable transduction with the p53 biosensors, we demonstrated that these orig...Continue Reading

References

Jan 27, 2000·Cell·D Hanahan, R A Weinberg
Nov 20, 2002·Proceedings of the National Academy of Sciences of the United States of America·R PaulmuruganS S Gambhir
Jun 21, 2003·Philosophical Transactions. Series A, Mathematical, Physical, and Engineering Sciences·Michele VendruscoloChristopher M Dobson
Dec 20, 2003·Nature·Christopher M Dobson
Jun 30, 2004·Trends in Endocrinology and Metabolism : TEM·Virginie BernierMichel Bouvier
Mar 1, 2005·Analytical Chemistry·Ramasamy Paulmurugan, Sanjiv S Gambhir
May 13, 2006·The FEBS Journal·Tapan K Chaudhuri, Subhankar Paul
Oct 18, 2006·Proceedings of the National Academy of Sciences of the United States of America·Ramasamy Paulmurugan, Sanjiv S Gambhir
Jan 4, 2008·Cancer Research·Carmel T ChanSanjiv Sam Gambhir
Feb 28, 2008·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Eszter Herczenik, Martijn F B G Gebbink
Dec 3, 2008·Molecular Imaging and Biology : MIB : the Official Publication of the Academy of Molecular Imaging·C T ChanS S Gambhir
Nov 26, 2009·Nature Reviews. Cancer·Christopher J BrownDavid P Lane
Feb 26, 2010·Cold Spring Harbor Perspectives in Biology·Magali OlivierPierre Hainaut
Jun 3, 2010·Cold Spring Harbor Perspectives in Biology·Andreas C Joerger, Alan R Fersht
Aug 17, 2012·Proceedings of the National Academy of Sciences of the United States of America·Carmel T ChanSanjiv S Gambhir
Apr 2, 2016·Nucleic Acids Research·Sinéad KearnsAndrei L Okorokov
Jun 10, 2016·Journal of Molecular Medicine : Official Organ of the Gesellschaft Deutscher Naturforscher Und Ärzte·Anjali V SheahanTarik F Massoud

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Citations

Aug 20, 2019·Oncology Letters·Fengyu ZhaiYi Zhang
Dec 11, 2019·International Journal of Molecular Sciences·Callum Hall, Patricia A J Muller
May 2, 2021·Biochimica Et Biophysica Acta. Reviews on Cancer·Liz J Hernández Borrero, Wafik S El-Deiry

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Methods Mentioned

BETA
ubiquitination
protein folding
PCAs
PCA
biosensors
biosensor
transfection
FACS
PCR
protein

Software Mentioned

ImageJ
Sequetech
FlowJo
Prism
Living Image quantitation
InCyte Software
Living Image
Guava
GraphPad

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