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Conference Paper: Sirtuin 1 interacts with DNA repair molecules during the initiation phase of reprogramming from mouse fibroblasts to mouse induced pluripotent stem cells
Title | Sirtuin 1 interacts with DNA repair molecules during the initiation phase of reprogramming from mouse fibroblasts to mouse induced pluripotent stem cells |
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Authors | |
Issue Date | 2014 |
Publisher | The International Society for Stem Cell Research (ISSCR). |
Citation | The 12th Annual Meeting of the International Society for Stem Cell Research (ISSCR 2014), Vancouver, Canada, 28-21 June 2014. In Conference Abstracts, 2014, p. 446, abstract no. F-2192 How to Cite? |
Abstract | The reprogramming of somatic cells to induced pluripotent stem cells
(iPSCs) involves chromatin remodeling. We have previously shown that
a NAD+ dependent histone deacetylase, sirtuin 1 (SIRT1) positively
regulated mouse iPSCs formation, partly through deacetylating p53
leading to altered Nanog and p21 expression (1). SIRT1 also deacetylates
other proteins like PGC1α [2] and FOXO3 [3]. The present study aimed
at identifying the SIRT1 interacting protein during the reprogramming
process. Pure population of secondary mouse embryonic fibroblast
(MEF) containing doxycycline (DOX) inducible OSKM factors were
obtained from chimeric MEF by Fluorescence Activated Cell Sorting.
The cells were cultured in the presence of DOX for the first 5 days
(initiation phase of reprogramming). The interacting proteins of SIRT1
during reprogramming were isolated by co-immunoprecipitation. The
immunoprecipitated proteins were visualized by silver staining, and
identified by mass spectrometry. According to the pathway clustering
analysis from the Kyoto Encyclopedia of Genes and Genomes
(KEGG), a number of them were involved in repair of double strand
break (DSB). Homologous recombination DNA repair genes are
important for successful reprogramming [4]. We speculated that
SIRT1 might promote DNA repair through binding to the complex of
DSB repair proteins. Indeed, the levels of the SIRT1 interacting DSB
repair proteins (SIRT1-DSB) were up-regulated in the MEF upon
DOX induction. To this end, we tried to knock down two of these
SIRT1-DSBs at the onset of reprogramming. The result showed that
the down-regulation of SIRT1-DSBs suppressed the iPSC colony
formation as demonstrated by lower colony number and reduced
Nanog expressions in the colonies. These data supported that SIRT1
activated the SIRT1-DSBs in the initial phase of reprogramming
process leading to increase in colony formation. Acknowledgement:
This work is supported by General Research Fund (HKU775711M)
and Seed Funding Scheme to Support Research Projects on
Human Embryonic Stem Cells (ESC) and Induced Pluripotent
Stem Cells (iPSC), Stem Cell and Regenerative Medicine
Consortium (SCRMC), Li Ka Shing Faculty of Medicine, The
University of Hong Kong. We thank Professor Andras Nagy (The
University of Toronto) for his generous gift of primary iPSCs. |
Description | Poster Session: iPS Cells |
Persistent Identifier | http://hdl.handle.net/10722/204889 |
DC Field | Value | Language |
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dc.contributor.author | Lee, CYL | en_US |
dc.contributor.author | Peng, Q | en_US |
dc.contributor.author | Chen, CH | en_US |
dc.contributor.author | Ng, EHY | en_US |
dc.contributor.author | Yeung, WSB | en_US |
dc.date.accessioned | 2014-09-20T00:56:01Z | - |
dc.date.available | 2014-09-20T00:56:01Z | - |
dc.date.issued | 2014 | en_US |
dc.identifier.citation | The 12th Annual Meeting of the International Society for Stem Cell Research (ISSCR 2014), Vancouver, Canada, 28-21 June 2014. In Conference Abstracts, 2014, p. 446, abstract no. F-2192 | en_US |
dc.identifier.uri | http://hdl.handle.net/10722/204889 | - |
dc.description | Poster Session: iPS Cells | - |
dc.description.abstract | The reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) involves chromatin remodeling. We have previously shown that a NAD+ dependent histone deacetylase, sirtuin 1 (SIRT1) positively regulated mouse iPSCs formation, partly through deacetylating p53 leading to altered Nanog and p21 expression (1). SIRT1 also deacetylates other proteins like PGC1α [2] and FOXO3 [3]. The present study aimed at identifying the SIRT1 interacting protein during the reprogramming process. Pure population of secondary mouse embryonic fibroblast (MEF) containing doxycycline (DOX) inducible OSKM factors were obtained from chimeric MEF by Fluorescence Activated Cell Sorting. The cells were cultured in the presence of DOX for the first 5 days (initiation phase of reprogramming). The interacting proteins of SIRT1 during reprogramming were isolated by co-immunoprecipitation. The immunoprecipitated proteins were visualized by silver staining, and identified by mass spectrometry. According to the pathway clustering analysis from the Kyoto Encyclopedia of Genes and Genomes (KEGG), a number of them were involved in repair of double strand break (DSB). Homologous recombination DNA repair genes are important for successful reprogramming [4]. We speculated that SIRT1 might promote DNA repair through binding to the complex of DSB repair proteins. Indeed, the levels of the SIRT1 interacting DSB repair proteins (SIRT1-DSB) were up-regulated in the MEF upon DOX induction. To this end, we tried to knock down two of these SIRT1-DSBs at the onset of reprogramming. The result showed that the down-regulation of SIRT1-DSBs suppressed the iPSC colony formation as demonstrated by lower colony number and reduced Nanog expressions in the colonies. These data supported that SIRT1 activated the SIRT1-DSBs in the initial phase of reprogramming process leading to increase in colony formation. Acknowledgement: This work is supported by General Research Fund (HKU775711M) and Seed Funding Scheme to Support Research Projects on Human Embryonic Stem Cells (ESC) and Induced Pluripotent Stem Cells (iPSC), Stem Cell and Regenerative Medicine Consortium (SCRMC), Li Ka Shing Faculty of Medicine, The University of Hong Kong. We thank Professor Andras Nagy (The University of Toronto) for his generous gift of primary iPSCs. | en_US |
dc.language | eng | en_US |
dc.publisher | The International Society for Stem Cell Research (ISSCR). | - |
dc.relation.ispartof | Annual Meeting of the International Society for Stem Cell Research, ISSCR 2014 | en_US |
dc.title | Sirtuin 1 interacts with DNA repair molecules during the initiation phase of reprogramming from mouse fibroblasts to mouse induced pluripotent stem cells | en_US |
dc.type | Conference_Paper | en_US |
dc.identifier.email | Lee, CYL: cherielee@hku.hk | en_US |
dc.identifier.email | Ng, EHY: nghye@hku.hk | en_US |
dc.identifier.email | Yeung, WSB: wsbyeung@hkucc.hku.hk | en_US |
dc.identifier.authority | Lee, CYL=rp00308 | en_US |
dc.identifier.authority | Ng, EHY=rp00426 | en_US |
dc.identifier.hkuros | 237526 | en_US |
dc.identifier.spage | 446, abstract no. F-2192 | - |
dc.identifier.epage | 446, abstract no. F-2192 | - |
dc.publisher.place | United States | - |