en
2008-2835
2008-4625
276
5669
11181
gregorian
>2015
>April-June
7
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online
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fulltext
en
26140185
TP53 Binding to BRCA1 and RAD51 in MCF7 and MDA-MB-468 Breast Cancer Cell Lines In vivo and In vitro
<p>Background: Tumour suppressor genes such as TP53, BRCA1 and RAD51 are involved in DNA repair and their malfunctions result in genomic instability and cancer. Wild type (WT) TP53 binds to BRCA1and RAD51 <em>in vivo</em> and<em> in vitro</em>. However, mutated TP53 in tumours can interfere with WT TP53 function. We studied how mutation of TP53 in MDA-MB-468 cell line could affect its binding capacity and interfere with WT TP53 interaction with these DNA repair proteins.<br />
Methods: Binding capacity of mutated TP53 in MDA-MB-468 breast cancer cell line to BRCA1 and RAD51 proteins in comparison to WT TP53 in MCF7 cell line was studied by Immunoprecipitation. <em>In vitro</em> studies were performed by GST-WT p53 pull-down assays in these cell lines to assess the interaction of GST-WT p53 with BRCA1 and RAD51 proteins.<br />
Results: The results showed that mutated TP53 in MDA-MB-468 cells interacted with BRCA1 protein<em> in vivo</em> and did not effect WT TP53 binding to this protein <em>in vitro</em>. The Immunoprecipitation assays revealed that the mutated TP53 did not bind to RAD51 in comparison to WT TP53. However, this mutated protein could not interfere with binding of RAD51 to GST-WT p53 in MDA-MB-468 cell line by in vitro experiment.<br />
Conclusion: It was found that WT TP53 interactions with BRCA1 and RAD51 did not interfere with mutated TP53 in MDA-MB-468 cell line. In addition, RAD51 did not bind to TP53 with R273C mutation<em> in vivo</em>.</p>
BRCA1 protein, Immunoprecipitation, RAD51 protein, TP53
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https://www.ajmb.org/En/Article.aspx?id=206
https://www.ajmb.org/PDF/En/FullText/206.pdf
MozhganRastiRecombinant Lab, Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran808
TayebehAzimiRecombinant Lab, Department of Biochemistry, Faculty of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran890