Knockdown of nucleosome assembly protein 1-like 1 promotes dimethyl sulfoxide-induced differentiation of P19CL6 cells into cardiomyocytes

2012 ◽  
Vol 113 (12) ◽  
pp. 3788-3796 ◽  
Author(s):  
Lu Li ◽  
Hui Gong ◽  
Hongxiu Yu ◽  
Xiaohui Liu ◽  
Qingping Liu ◽  
...  
Keyword(s):  
Dimethyl Sulfoxide ◽  
Nucleosome Assembly ◽  
Induced Differentiation ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1 ◽  
P19cl6 Cells

Abstract 193: Knockdown of Nucleosome Assembly Protein 1-Like 1 Promotes Dimethyl Sulfoxide-Induced Differentiation of P19CL6 Cells into Cardiomyocytes

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Hui Gong ◽  
Lu Li ◽  
Yuan Yan ◽  
Yuanyuan Xue ◽  
Zhidan Chen ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Therapy ◽  
Dimethyl Sulfoxide ◽  
Real Time ◽  
Stem Cell Therapy ◽  
Stem Cell Differentiation ◽  
Nucleosome Assembly ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1 ◽  
P19cl6 Cells

Various stem cell populations have the potential for cardiac repair and regeneration after transplanted into ischaemic injury heart. However, the low generation of new cardiomyocytes limits the clinic outcomes of stem cell therapy to a great extent. To develop novel strategies for enhancement the efficacy of stem cell differentiation into cardiomyocytes, we employed iTRAQ-coupled 2-D LC-MS/MS proteomics technology workflow to identify novel proteins which could promote P19CL6 cells differentiation into cardiomyocytes induced by dimethyl sulfoxide dimethyl sulfoxide (DMSO). 213 differentially expressed proteins were identified during differentiation of P19CL6 cells into cardiomyocytes. Nine proteins, Nap1L1, Sub1, Hspa5, Ywhaz, Sod1, Prdx2, Mif, Anp32a and Eef2, were confirmed by the real-time polymerase chain reaction (PCR) and western blotting respectively. iTRAQ ratios showed the down-regulation of nucleosome assembly protein 1-like 1 (Nap1L1) during the differentiation of P19CL6 cells into cardiomyocytes, which was confirmed by real-time PCR and western blot results. Further study indicated that knockdown of Nap1L1 expression promoted DMSO-induced cardiomyocytes differentiation in P19CL6 cells characterized by the increased expresison of cardiac specilal transcription factors (GATA4, MEF2C) and proteins (cTNT, ANP and BNP). This work suggested that Nap1L1 play an important role in DMSO-induced cardiomyocytes differentiation of P19CL6 cells. It also provides new insights and platforms for stem cell therapy in cardiovascular disease.

Abstract 140: Knockdown of Nucleosome Assembly Protein 1-like 1 Promotes Cardiomyocytes Differentiation by Mesoderm induction through Notch Signaling in Mouse Induced Pluripotent Stem Cells

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
Hui Gong ◽  
Yuan Yan ◽  
Yuanyuan Xue ◽  
Peipei Yin ◽  
Guoping Zhang ◽  
...  
Keyword(s):  
Notch Signaling ◽  
Heart Diseases ◽  
Es Cells ◽  
Mesoderm Induction ◽  
Nucleosome Assembly ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1 ◽  
P19cl6 Cells ◽  
Induced Pluripotent ◽  
Cardiac Transcription Factors

Recently, we used a functional proteomic analysis to screen out nucleosome assembly protein 1-like 1 (Nap1l1) which was downregulated during the differentiation of P19CL6 cells into cardiomyocytes. Here, we attempted to study the role of Nap1l1 in the cardiogenesis of mouse iPSCs. We observed Nap1l1 was downregulated during the differentiation of iPSCs. Knockdown of Nap1l1 dramatically enhanced the differentiation of iPSCs to cardiomyocytes characterized by the increased number of beating embryonic bodies (EBs), the larger alpha-myosin heavy chain (α-MHC)-stained area and the upregulation of cardiac transcription factors (Nkx2.5, GATA4, Mef2c, Tbx5). The effects were sharply inhibited by Nap1l1 overexpression in iPSCs. Cardiomyocytes derived from Nap1l1-knockdown-iPSCs exhibited proper cell biological characteristics. Further study revealed that Nap1l1 knockdown in iPSCs promoted mesoderm (Flk-1, Brachyury and Mesp1) development, but Nap1l1 overexpression inhibited the effect. To explore whether Nap1l1 knockdown in iPSCs enhances cardiomyocytes differentiation by mesoderm induction. Mesoderm cells (Flk-1 positive cells) from iPSCs development were sorted by fluorescent-assisted cell sorting (FACS) and recultured to induce cardiomyocytes differentiation. The result revealed that the same number of Flk-1(mesodermal marker) positive cells from Nap1l1 knockdown, Nap1l1 overexpression or their control iPSCs didn’t show obvious difference in cardiomyocyte differentiation. Loss of Notch signaling in ES cells has been reported to favor commitment to a mesoderm and to induce cardiogenesis. The present study revealed that NICD and downstream genes (Hes1, Hes5, Hey1 and Hey 2) were positively regulated by Nap1l1 expression during differentiation of iPSCs. Notch signaling inhibitor greatly rescued the inhibitory effects of Nap1l1 overexpression on mesoderm induction and cardiogenesis. These findings demonstrate that downregulation of Nap1l1 significantly enhances mesodermal induction and subsequently promotes cardiogenesis from mouse iPSCs via regulating Notch signaling, which will facilitate application of iPSCs for heart diseases.

Assembly states of the nucleosome assembly protein 1 (NAP-1) revealed by sedimentation velocity and non-denaturing MS

2011 ◽  
Vol 436 (1) ◽  
pp. 101-112 ◽  
Author(s):  
Masanori Noda ◽  
Susumu Uchiyama ◽  
Adam R. McKay ◽  
Akihiro Morimoto ◽  
Shigeki Misawa ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Electrostatic Interactions ◽  
Analytical Ultracentrifugation ◽  
Three Dimensional ◽  
Sedimentation Velocity ◽  
Dimensional Structure ◽  
Histone H2a ◽  
Nucleosome Assembly ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1

Proteins often exist as ensembles of interconverting states in solution which are often difficult to quantify. In the present manuscript we show that the combination of MS under nondenaturing conditions and AUC-SV (analytical ultracentrifugation sedimentation velocity) unambiguously clarifies a distribution of states and hydrodynamic shapes of assembled oligomers for the NAP-1 (nucleosome assembly protein 1). MS established the number of associated units, which was utilized as input for the numerical analysis of AUC-SV profiles. The AUC-SV analysis revealed that less than 1% of NAP-1 monomer exists at the micromolar concentration range and that the basic assembly unit consists of dimers of yeast or human NAP-1. These dimers interact non-covalently to form even-numbered higher-assembly states, such as tetramers, hexamers, octamers and decamers. MS and AUC-SV consistently showed that the formation of the higher oligomers was suppressed with increasing ionic strength, implicating electrostatic interactions in the formation of higher oligomers. The hydrodynamic shapes of the NAP-1 tetramer estimated from AUC-SV agreed with the previously proposed assembly models built using the known three-dimensional structure of yeast NAP-1. Those of the hexamer and octamer could be represented by new models shown in the present study. Additionally, MS was used to measure the stoichiometry of the interaction between the human NAP-1 dimer and the histone H2A–H2B dimer or H3–H4 tetramer. The present study illustrates a rigorous procedure for the analysis of protein assembly and protein–protein interactions in solution.

Organization and expression of the Paramecium caudatum gene encoding nucleosome assembly protein 1

Gene ◽  
2001 ◽  
Vol 280 (1-2) ◽  
pp. 107-114 ◽  
Author(s):  
Norihito Nishiyama ◽  
Shun Sawatsubashi ◽  
Masaki Ishida ◽  
Kiyoshi Yamauchi
Keyword(s):  
Paramecium Caudatum ◽  
Nucleosome Assembly ◽  
Gene Encoding ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1

Self-Association of the Yeast Nucleosome Assembly Protein 1

Biochemistry ◽  
2004 ◽  
Vol 43 (32) ◽  
pp. 10592-10599 ◽  
Author(s):  
Steven J. McBryant ◽  
Olve B. Peersen
Keyword(s):  
Nucleosome Assembly ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1 ◽  
Self Association

Biochemical and Biophysical Characterisation of Higher Oligomeric Structure of Rat Nucleosome Assembly Protein 1

2017 ◽  
Vol 37 (1) ◽  
pp. 58-69
Author(s):  
Divya Reddy ◽  
Saikat Bhattacharya ◽  
Vinod Jani ◽  
Uddhavesh Sonavane ◽  
Rajendra Joshi ◽  
...  
Keyword(s):  
Nucleosome Assembly ◽  
Oligomeric Structure ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1

scholarly journals Nucleosome Assembly Protein 1 Exchanges Histone H2A-H2B Dimers and Assists Nucleosome Sliding

2004 ◽  
Vol 280 (3) ◽  
pp. 1817-1825 ◽  
Author(s):  
Young-Jun Park ◽  
Jayanth V. Chodaparambil ◽  
Yunhe Bao ◽  
Steven J. McBryant ◽  
Karolin Luger
Keyword(s):  
Histone H2a ◽  
Nucleosome Assembly ◽  
Nucleosome Assembly Protein ◽  
Nucleosome Assembly Protein 1 ◽  
Nucleosome Sliding
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