Mutational Analysis of Progesterone Receptor Functional Domains in Stable Cell Lines Delineates Sets of Genes Regulated by Different Mechanisms
| Autor: | Nora Spinedi, Belén Miñana, Ignacio Quiles, Albert Jordan, Alicia Subtil-Rodríguez, Miguel Beato, Cecilia Ballaré, Lluís Millán-Ariño |
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| Jazyk: | angličtina |
| Rok vydání: | 2009 |
| Předmět: |
Gene isoform
MAPK/ERK pathway Time Factors DNA Mutational Analysis Estrogen receptor Biology Transactivation Endocrinology Cell Line Tumor Progesterone receptor Humans Cyclin D1 RNA Messenger Promoter Regions Genetic Molecular Biology Estrogen receptor beta Original Research Cell Proliferation Oligonucleotide Array Sequence Analysis Regulation of gene expression Models Genetic Reverse Transcriptase Polymerase Chain Reaction Gene Expression Profiling General Medicine Molecular biology Protein Structure Tertiary Enzyme Activation Gene Expression Regulation Neoplastic Kinetics Mutant Proteins Signal transduction Progestins Receptors Progesterone Protein Kinases Protein Binding Signal Transduction |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | Steroid hormone receptors act directly in the nucleus on the chromatin organization and transcriptional activity of several promoters. Furthermore, they have an indirect effect on cytoplasmic signal transduction pathways, including MAPK, impacting ultimately on gene expression. We are interested in distinguishing between the two modes of action of progesterone receptor (PR) on the control of gene expression and cell proliferation. For this, we have stably expressed, in PR-negative breast cancer cells, tagged forms of the PR isoform B mutated at regions involved either in DNA binding (DNA-binding domain) or in its ability to interact with the estrogen receptor and to activate the c-Src/MAPK/Erk/Msk cascade (estrogen receptor-interacting domain). Both mutants impair PR-mediated activation of a well-understood model promoter in response to progestin, as well as hormone-induced cell proliferation. Additional mutants affecting transactivation activity of PR (activation function 2) or a zinc-finger implicated in dimerization (D-box) have also been tested. Microarrays and gene expression experiments on these cell lines define the subsets of hormone-responsive genes regulated by different modes of action of PR isoform B, as well as genes in which the nuclear and nongenomic pathways cooperate. Correlation between CCND1 expression in the different cell lines and their ability to support cell proliferation confirms CCND1 as a key controller gene. Copyright © 2009 by The Endocrine Society. This work was supported by grants from the Catalan Department for Universities, Research and the Information Society, and the Spanish Ministry of Science and Technology and Fondo Europeo de desarrollo regional (SAF2002-03320, BFU2008-00359). A.J. was recipient of a ‘Ramón y Cajal’ appointment from the Spanish Ministry of Science and Technology. I.Q. was recipient of a Formación personal universitario predoctoral fellowship from the Spanish Ministry of Education. L.M-A. was recipient of a predoctoral fellowship funded by Fundación para la investigación y prevención del SIDA en España |
| Databáze: | OpenAIRE |
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