Dao, K.-H.T., Rotelli, M.D., Brown, B.R. et al. (6 more authors) (2013) The PI3K/Akt1 pathway enhances steady-state levels of FANCL. Molecular Biology of the Cell, 24 (16). pp. 2582-2592.
Abstract
Fanconi anemia hematopoietic stem cells display poor self-renewal capacity when subjected to a variety of cellular stress. This phenotype raises the question of whether the Fanconi anemia proteins are stabilized or recruited as part of a stress response and protect against stem cell loss. Here we provide evidence that FANCL, the E3 ubiquitin ligase of the Fanconi anemia pathway, is constitutively targeted for degradation by the proteasome. We confirm biochemically that FANCL is polyubiquitinated with Lys-48-linked chains. Evaluation of a series of N-terminal-deletion mutants showed that FANCL's E2-like fold may direct ubiquitination. In addition, our studies showed that FANCL is stabilized in a complex with axin1 when glycogen synthase kinase-3β is overexpressed. This result leads us to investigate the potential regulation of FANCL by upstream signaling pathways known to regulate glycogen synthase kinase-3β. We report that constitutively active, myristoylated-Akt increases FANCL protein level by reducing polyubiquitination of FANCL. Two-dimensional PAGE analysis shows that acidic forms of FANCL, some of which are phospho-FANCL, are not subject to polyubiquitination. These results indicate that a signal transduction pathway involved in self-renewal and survival of hematopoietic stem cells also functions to stabilize FANCL and suggests that FANCL participates directly in support of stem cell function.
Metadata
Item Type: | Article |
---|---|
Authors/Creators: |
|
Copyright, Publisher and Additional Information: | © 2013 Dao et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). |
Keywords: | Axin Protein; Cell Line; Enzyme Activation; Fanconi Anemia; Fanconi Anemia Complementation Group L Protein; Gene Expression; Glycogen Synthase Kinase 3; Glycogen Synthase Kinase 3 beta; HEK293 Cells; HeLa Cells; Hematopoietic Stem Cells; Humans; Phosphatidylinositol 3-Kinases; Proteasome Endopeptidase Complex; Protein Folding; Proto-Oncogene Proteins c-akt; RNA Interference; RNA, Small Interfering; Signal Transduction; Ubiquitination |
Dates: |
|
Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 15 Nov 2019 12:04 |
Last Modified: | 15 Nov 2019 15:28 |
Status: | Published |
Publisher: | American Society for Cell Biology |
Refereed: | Yes |
Identification Number: | 10.1091/mbc.E13-03-0144 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:153365 |
Download
Filename: The PI3KAkt1 pathway enhances steady-state levels of FANCL.pdf
Licence: CC-BY-NC-SA 3.0