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Functional Study of a New Protein ERIP1 Interacting with ER

Author: JiangYanChao
Tutor: HuJianMin;YeQiNong
School: Shenyang Agricultural University
Course: Basic Veterinary
Keywords: ERIP1 ER Protein interactions Signal transduction
CLC: Q51
Type: Master's thesis
Year: 2007
Downloads: 23
Quote: 0
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Executive Summary


Estrogen receptor ER is a nuclear receptor, regulate estrogen-related gene transcription, has an important role in the development process in the incidence of breast cancer, breast cancer treatment target and prognosis. ER is divided into ERα and ERβ two kinds, both of which are A, B, C, D, E and F, six structural domains, containing the two transcriptional activation AF1 and AF2. AF1 with estrogen-dependent transcriptional activation function located in the A / B region; AF2 with estrogen-dependent transcriptional activation function located in the E / F region. AF1 and AF2 efficient transcription activation depends on the interaction with other proteins. The discovery and function of proteins involved in regulating ER transcriptional activity of great significance for the effective development of drugs to treat breast cancer. ERIP1 an unknown protein, has not yet been reflected in its research reports. This paper is a preliminary study on the basis of the yeast two-hybrid ERIP1 to determine the interactions between ERIP1 with ER and ERIP1 ER downstream reporter gene activity, and as a preliminary exploration of the biological function of ERIP1 . In this study, the PCR method successfully from the mammary gland cDNA library tune to take a ERIP1 gene to its building to the tests required for the original nuclear eukaryotic expression vector, these recombinant plasmid corresponding original nuclear, true nuclear cells have been correctly expression; by GST pull-down experiments show, ERIP1 in vitro can be combined and ER binding and ERα ERIP1 of a, b, and c fragment, wherein b fragment of the strongest binding capacity, its strength over the entire length, and ERβ binding ERIP1 of b, the C fragment; intracellular co-immunoprecipitation experiments show that, in a hormone dependent manner ERIP1 in vivo interaction with ERβ, ERIP1 four fragments can be combined with ER, wherein b fragment with ERIP1 ERα binding capacity of the strongest. Comprehensive three results can be used to validate the protein-protein interaction experiments concluded existence: ERIP1 with ER interaction. In order to explore the biological functions of ERIP1, we conducted a series of active experiments. ERE-luc, pS2-luc C3-luc-luc, CatD differ, ERIP1 ER downstream reporter gene, the same trend of ERIP1 the ERE-luc, pS2-luc and elevated ERα transcriptional activity, reduced the transcriptional activity of ERβ. Showed ERIP1 two ER binding role is to promote the occurrence and development of cancer in breast cancer, the role of ERα and ERβ opposite. ERIP1 were co-transfected with ER C3-luc activity. ERIP1 with ER can collaborate with elevated CatD-luc activity rpm single elevated CatD-luc activity, the transcription factor through transcriptional activity proved ERIP1 not, it not only as a ER co-regulatory factors affecting CatD- luc activity, also may interact with certain other factors change CatD-luc activity through another pathway, and a dose-dependent effect. In immunohistochemistry experiments, with the GFP label ERIP1 transfected into HEK-293T and MCF-7 cells, found that the death of cells in a large area transit cells transfected into a plasmid, nuclear shrinkage, crushing, the instructions ERIP1 may induce apoptosis. These results show that, ERIP1 may be ER signaling pathways in a new co-regulatory factors, thus affecting the interaction with ER ER downstream signal transfer, further in-depth study will help to understand the regulatory mechanism of the ER signaling pathway ERIP1 clinical therapeutic effect, it is possible to provide a new target for the treatment of breast cancer.

Full-text Catalog


Chinese Abstract     6-8
English summary     8-9
Preface     9-19
the
the first chapter ERIP1 with ER interaction     19-35
1 Materials and methods     19-27
2 Results and Analysis     27-33
3 Discussion     33-34
4 Summary     34-35
Chapter ERIP1 . ER transcriptional activity     35-45
1 Materials and methods     35-36
2 Results and Analysis     36-42
3 Discussion     42-44
4 Summary     44-45
the
ERIP1 apoptosis     45-47
1 Materials and methods     45
2 Results and Analysis     45-46
3 Discussion     46
4 Summary     46-47
concluded     47-48
the
References     48-53
Appendix     53-56
Acknowledgements     56-57
the
studying dissertation published during the     57

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