دورية أكاديمية
Pim-1 kinase inhibits the activation of reporter gene expression in Elk-1 and c-Fos reporting systems but not the endogenous gene expression: an artifact of the reporter gene assay by transient co-transfection
العنوان: | Pim-1 kinase inhibits the activation of reporter gene expression in Elk-1 and c-Fos reporting systems but not the endogenous gene expression: an artifact of the reporter gene assay by transient co-transfection |
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المؤلفون: | B. Yan, H. Wang, T. Kon, C.-Y. Li |
المصدر: | Brazilian Journal of Medical and Biological Research, Vol 39, Iss 2, Pp 169-176 (2006) |
بيانات النشر: | Associação Brasileira de Divulgação Científica, 2006. |
سنة النشر: | 2006 |
المجموعة: | LCC:Medicine (General) LCC:Biology (General) |
مصطلحات موضوعية: | Pim-1 kinase, Signal transduction, Mitogen-activated protein kinase pathway, Elk-1, Reporter assay, Medicine (General), R5-920, Biology (General), QH301-705.5 |
الوصف: | We have studied the molecular mechanism and signal transduction of pim-1, an oncogene encoding a serine-threonine kinase. This is a true oncogene which prolongs survival and inhibits apoptosis of hematopoietic cells. In order to determine whether the effects of Pim-1 occur by regulation of the mitogen-activated protein kinase pathway, we used a transcriptional reporter assay by transient co-transfection as a screening method. In this study, we found that Pim-1 inhibited the Elk-1 and NFkappaB transcriptional activities induced by activation of the mitogen-activated protein kinase cascade in reporter gene assays. However, Western blots showed that the induction of Elk-1-regulated expression of endogenous c-Fos was not affected by Pim-1. The phosphorylation and activation of neither Erk1/2 nor Elk-1 was influenced by Pim-1. Also, in the gel shift assay, the pattern of endogenous NFkappaB binding to its probe was not changed in any manner by Pim-1. These data indicate that Pim-1 does not regulate the activation of Erk1/2, Elk-1 or NFkappaB. These contrasting results suggest a pitfall of the transient co-transfection reporter assay in analyzing the regulation of transcription factors outside of the chromosome context. It ensures that results from reporter gene expression assay should be verified by study of endogenous gene expression. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English |
تدمد: | 1414-431X 0100-879X |
Relation: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2006000200002&lng=en&tlng=en; https://doaj.org/toc/1414-431X |
DOI: | 10.1590/S0100-879X2006000200002 |
URL الوصول: | https://doaj.org/article/844e80f01c4e407896a4b6d12a93b6a9 |
رقم الأكسشن: | edsdoj.844e80f01c4e407896a4b6d12a93b6a9 |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 1414431X 0100879X |
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DOI: | 10.1590/S0100-879X2006000200002 |