Multiple post-translational modifications in hepatocyte nuclear factor 4α

Yokoyama, Atsushi; Katsura, Shogo; Itô, Ryo; Hashiba, Waka; Sekine, Hiroshi; Fujiki, Ryoji; Kato, Shigeaki

doi:10.1016/j.bbrc.2011.06.033

Cited by 38 publications

(49 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In-gel digestion of sperm histones was performed as described previously, with some modifications46. The sperm S2 fraction was resolved by Any kD Mini-PROTEAN TGX Precast Protein Gels (Bio-Rad, Hercules, CA, USA), then visualized using a Silver Stain Kit (Bio-Rad).…”

Section: Methodsmentioning

confidence: 99%

Identification of a variant-specific phosphorylation of TH2A during spermiogenesis

Hada

Masuda

Yamaguchi

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Tissue-specific histone variant incorporation into chromatin plays dynamic and important roles in tissue development. Testis is one such tissue, and a number of testis-specific histone variants are expressed that have unique roles. While it is expected that such variants acquire post-transcriptional modifications to be functional, identification of variant-specific histone modifications is challenging because of the high similarity of amino acid sequences between canonical and variant versions. Here we identified a novel phosphorylation on TH2A, a germ cell-specific histone H2A variant. TH2A-Thr127 is unique to the variant and phosphorylated concomitant with chromatin condensation including spermiogenesis and early embryonic mitosis. In sperm chromatin, phosphorylated TH2A-Thr127 (=pTH2A) is co-localized with H3.3 at transcriptional starting sites of the genome, and subsequently becomes absent from the paternal genome upon fertilization. Notably, pTH2A is recurrent and accumulated in the pericentromeric heterochromatin of both paternal and maternal chromosomes in the first mitosis of embryos, suggesting its unique regulation during spermiogenesis and early embryogenesis.

show abstract

Section: Methodsmentioning

confidence: 99%

Identification of a variant-specific phosphorylation of TH2A during spermiogenesis

Hada

Masuda

Yamaguchi

et al. 2017

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Purification of PPAR γ protein was performed as previously described [16]. Briefly, we cross-linked 2 μ g of antibody with 30 μ L of Protein G Dynabeads (Invitrogen) in freshly dissolved 20 mM dimethyl pimelimidate (DMP) in a 0.2 M triethanolamine buffer (pH 8.2).…”

Section: Methodsmentioning

confidence: 99%

“…Mass spectrometric (MS) analysis of PPAR γ protein and PTM analysis were performed as previously described [16]. Briefly, PPAR γ protein was excised from the gel, reduced with 10 mM dithiothreitol solution in 0.1 M ammonium bicarbonate for 60 min at 56°C, alkylated with a 55 mM solution of iodoacetamide in 0.1 M ammonium bicarbonate in darkness for 45 min at room temperature, and in-gel digested with 25 ng/ μ L trypsin gold (Promega, Madison, WI) in 50 mM ammonium bicarbonate for 16 h at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…Protein identification and PTM analysis of PPAR γ protein were performed as previously described [16]. Briefly, for protein identification, spectra were processed using Proteome Discoverer ver.…”

Section: Methodsmentioning

confidence: 99%

“…Protein posttranslational modifications (PTMs) are one of the most efficient biological signals for expanding the genetic code and play key roles in diverse cellular processes such as protein transportation, DNA repair, and gene transcription [13–15]. We have previously developed a comprehensive PTM analysis system for proteins, combining biochemical and proteomic approaches [16]. In this system, we utilized nonrestrictive sequence alignment for PTM analysis and this method made it possible to identify PTMs without prior specification, enabling unbiased analysis of PTMs [17, 18].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Identification of Posttranslational Modifications in Peroxisome Proliferator-Activated ReceptorγUsing Mass Spectrometry

et al. 2014

Self Cite

View full text Add to dashboard Cite

Posttranslational modification (PTM) of proteins is critical for various cellular processes. However, there are few studies examining PTMs in specific proteins using unbiased approaches. Here we report the attempt to identify the PTMs in peroxisome proliferator-activated receptor γ (PPARγ) proteins using our previously established PTM analysis system. In this study, we identified several PTMs in exogenously expressed PPARγ2 proteins from 293T cells as well as endogenous PPARγ1 proteins from a Caco-2 colon cancer cell line. The identified PTMs include phosphorylation of serine 112 and serine 81 in PPARγ2 and PPARγ1, respectively, both of which are well-known mitogen-activated protein kinase- (MAP kinase-) mediated PTMs in PPARγ proteins, thus confirming our experimental approach. Furthermore, previously unknown PTMs were also identified, demonstrating that this method can be applied to find previously unidentified PTMs in PPARγ proteins and other proteins including nuclear receptors.

show abstract

Hepatocyte Nuclear Factor 4 Alpha Activation Is Essential for Termination of Liver Regeneration in Mice

et al. 2019

View full text Add to dashboard Cite

Hepatocyte nuclear factor 4 alpha (HNF4α) is critical for hepatic differentiation. Recent studies have highlighted its role in inhibition of hepatocyte proliferation and tumor suppression. However, the role of HNF4α in liver regeneration (LR) is not known. We hypothesized that hepatocytes modulate HNF4α activity when navigating between differentiated and proliferative states during LR. Western blotting analysis revealed a rapid decline in nuclear and cytoplasmic HNF4α protein levels, accompanied with decreased target gene expression, within 1 hour after two‐thirds partial hepatectomy (post‐PH) in C57BL/6J mice. HNF4α protein expression did not recover to pre‐PH levels until day 3. Hepatocyte‐specific deletion of HNF4α (HNF4α‐KO [knockout]) in mice resulted in 100% mortality post‐PH, despite increased proliferative marker expression throughout regeneration. Sustained loss of HNF4α target gene expression throughout regeneration indicated that HNF4α‐KO mice were unable to compensate for loss of HNF4α transcriptional activity. Deletion of HNF4α resulted in sustained proliferation accompanied by c‐Myc and cyclin D1 overexpression and a complete deficiency of hepatocyte function after PH. Interestingly, overexpression of degradation‐resistant HNF4α in hepatocytes delayed, but did not prevent, initiation of regeneration after PH. Finally, adeno‐associated virus serotype 8 (AAV8)‐mediated reexpression of HNF4α in hepatocytes of HNF4α‐KO mice post‐PH restored HNF4α protein levels, induced target gene expression, and improved survival of HNF4α‐KO mice post‐PH. Conclusion: In conclusion, these data indicate that HNF4α reexpression following initial decrease is critical for hepatocytes to exit from cell cycle and resume function during the termination phase of LR. These results indicate the role of HNF4α in LR and have implications for therapy of liver failure.

show abstract

Multiple post-translational modifications in hepatocyte nuclear factor 4α

Cited by 38 publications

References 35 publications

Identification of a variant-specific phosphorylation of TH2A during spermiogenesis

Identification of a variant-specific phosphorylation of TH2A during spermiogenesis

Identification of Posttranslational Modifications in Peroxisome Proliferator-Activated ReceptorγUsing Mass Spectrometry

Hepatocyte Nuclear Factor 4 Alpha Activation Is Essential for Termination of Liver Regeneration in Mice

Contact Info

Product

Resources

About