scholarly journals Progesterone-induced Warburg Effect is Regulated by Cell-type-specific Interaction of Progesterone Receptor Membrane Component 1 and Hexokinases

2020 ◽  
Author(s):  
Mohammad Golam Sabbir ◽  
Carla G Taylor ◽  
Peter Zahradka
Keyword(s):  
Glucose Metabolism ◽  
Progesterone Receptor ◽  
Hepg2 Cells ◽  
Specific Interaction ◽  
Metabolic Reprogramming ◽  
Hek293 Cells ◽  
Cell Type ◽  
Membrane Component ◽  
Cell Type Specific ◽  
Hormone Responsiveness

Abstract Background: Progesterone receptor membrane component 1 (PGRMC1) is a non-canonical progesterone (P4) binding protein. PGRMC1 is elevated in a variety of cancers and its phosphorylation state associated with hormone responsiveness in breast cancer. Metabolic reprogramming is a key factor for tumor growth during malignancies. Recently, we reported that the P4-inducedWarburg effectinHEK293cells is associated with altered post-translational modifications (PTMs) of PGRMC1, including phosphorylation, SUMOylation, and ubiquitination, which were linked to rapid proteasomal degradation of the protein. The previous study also identified hexokinase (HK) as a potential novel interacting partner of PGRMC1. HKs catalyze the first essential step of glucose metabolism and directly couple glycolysis to mitochondrial respiration. Therefore, in the present study, P4’s effects on glycolysis and PTMs of PGRMC1 as well as its interaction with HKs were compared between HEK293 and HepG2 cells to unravel the signaling pathways that mediate cell-type-specific metabolic reprogramming.Methods: P4-induced glucose metabolism in wild-type and PGRMC1-deficient cells wasassessed using the Seahorse flux analyzer, while PTMs of PGRMC1/HKs and protein-protein interaction were studied using immunoprecipitation, isoelectric focussing, phosphomimetics, and mass spectrometry.Translocation of HKs to different subcellular organelles were studied using subcellular fractionation, and the cell-type-specific effect of PGRMC1-deficiency on endoplasmic reticulum (ER) and mitochondria; ultrastructure were examined by electron microscopy. Results:P4 treatment caused a rapid increase in glycolysis in HEK293 cells, whereas it decreased glycolysis in HepG2 cells. In addition, PGRMC1 was not degraded in HepG2 cells which is in contrast to HEK293 cells where rapid proteasomal degradation of PGRMC1 occurredfollowing P4 treatment. Besides, PGRMC1 half-life and PTMs under basal condition were found cell-type-specific and the P4-induced PTMsdiffered between the two cell types. Furthermore, we observed cell-type-specific interaction of HKs with PGRMC1, and differential translocation of HK1/2 to the ER, mitochondria and nuclear compartments following P4 treatment. PGRMC1 deficiency altered ER structure in HepG2 cells. Thus, multiple factors underlying the cell-type-specific P4-PGRMC1-mediated metabolic reprogrammingwere identified. Conclusions: These findings provide a hitherto unknown novel P4-induced cell-type-specific PGRMC1-HK signaling mechanism that contributes to the molecular basis of P4-induced metabolic reprogramming, with important applications for hormone responsiveness in cancer.

scholarly journals Cell Type-specific Target Selection by Combinatorial Binding of Smad2/3 Proteins and Hepatocyte Nuclear Factor 4α in HepG2 Cells

2011 ◽  
Vol 286 (34) ◽  
pp. 29848-29860 ◽  
Author(s):  
Anna Mizutani ◽  
Daizo Koinuma ◽  
Shuichi Tsutsumi ◽  
Naoko Kamimura ◽  
Masato Morikawa ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Target Selection ◽  
Nuclear Factor ◽  
Hepatocyte Nuclear Factor ◽  
Cell Type ◽  
Specific Target ◽  
Cell Type Specific ◽  
Hepatocyte Nuclear Factor 4Α

Cell-Type Specific Actions of Progesterone Receptor Modulators in the Regulation of Uterine Leiomyoma Growth

2010 ◽  
Vol 28 (03) ◽  
pp. 260-273 ◽  
Author(s):  
Shigeki Yoshida ◽  
Noriyuki Ohara ◽  
Qin Xu ◽  
Wei Chen ◽  
Jiayin Wang ◽  
...  
Keyword(s):  
Progesterone Receptor ◽  
Uterine Leiomyoma ◽  
Cell Type ◽  
Cell Type Specific ◽  
Receptor Modulators ◽  
Progesterone Receptor Modulators

scholarly journals Proteolytic processing and glycosylation of cathepsin B. The role of the primary structure of the latent precursor and of the carbohydrate moiety for cell-type-specific molecular forms of the enzyme

1992 ◽  
Vol 282 (2) ◽  
pp. 577-582 ◽  
Author(s):  
L Mach ◽  
K Stüwe ◽  
A Hagen ◽  
C Ballaun ◽  
J Glössl
Keyword(s):  
Heavy Chain ◽  
Cathepsin B ◽  
Hepg2 Cells ◽  
Cysteine Proteinase ◽  
Carbohydrate Moiety ◽  
Molecular Forms ◽  
Cell Type ◽  
Single Chain ◽  
Cell Type Specific ◽  
Chain Form

The lysosomal cysteine proteinase cathepsin B is synthesized in cultured human hepatoma HepG2 cells as an inactive 44 kDa precursor and subsequently processed to the mature single-chain enzyme with a molecular mass of 33 kDa. Intralysosomal conversion into the two-chain form results in subunits of 27 kDa, 24 kDa (heavy chain) and 5 kDa (light chain). Enzymic deglycosylation reveals that the 27 kDa polypeptide is the glycosylated variant of the carbohydrate-free 24 kDa heavy-chain form. The intracellular transport to the lysosomes is dependent upon mannose 6-phosphate-containing N-linked oligosaccharides. Receptor-mediated endocytosis of human skin-fibroblast-derived procathepsin B by HepG2 cells resulted in processed molecular forms that are not distinguishable from endogenous cathepsin B, thus favouring rather a cell-type-specific processing than structural differences due to the source of the proenzyme. The conversion step of single-chain catehpsin B into the two-chain enzyme is inhibited in vivo by the irreversible cysteine-proteinase inhibitors Z-Phe-Ala-CHN2 and, albeit weaker, Z-Phe-Phe-CHN2. Both substances have no effect on the activation of procathepsin B to the mature enzyme. The carbohydrate moiety of cathepsin B exerts no significant influence on the stability and the enzymatic activity of the enzyme.

scholarly journals Interaction studies of risk proteins in human induced neurons reveal convergent biology and novel mechanisms underlying autism spectrum disorders

2021 ◽  
Author(s):  
Greta Pintacuda ◽  
Yu-Han H Hsu ◽  
Kalliopi Tsafou ◽  
Ka Wan Li ◽  
Jacqueline M Martin ◽  
...  
Keyword(s):  
Autism Spectrum Disorders ◽  
Rare Variant ◽  
Specific Interaction ◽  
Autism Spectrum ◽  
Brain Cell ◽  
Synaptic Proteins ◽  
Cell Type ◽  
Risk Genes ◽  
Cell Type Specific ◽  
Spectrum Disorders

Sequencing studies of autism spectrum disorders (ASDs) have identified numerous risk genes with enriched expression in the human brain, but it is still unclear how these genes converge into cell type-specific networks and how their encoded proteins mechanistically contribute to ASDs. To address this question, we performed brain cell type-specific interaction proteomics to build a protein-protein interaction network for 13 ASD risk genes in human excitatory neurons derived from iPS cells. The network contains many (>90%) reproducible interactions not reported in the literature and is enriched for transcriptionally perturbed genes observed in layer 2/3 cortical neurons of ASD patients, indicating that it can be explored for ASD-relevant biological discovery. We leveraged the network dataset to show that the brain-specific isoform of ANK2 is important for its interactions with synaptic proteins and characterized a PTEN-AKAP8L interaction that influences neuronal growth through the mTOR pathway. The IGF2BP1-3 complex emerges as a point of convergence in the network, and we showed that this complex is involved in a transcriptional circuit concentrating both common and rare variant risk of ASDs. Finally, we found the network itself enriched for ASD rare variant risk, indicating that it can complement genetic datasets for prioritizing additional risk genes. Our findings establish brain cell type-specific interactomes as an organizing framework to facilitate interpretation of genetic and transcriptomic data in ASDs and illustrate how both individual and convergent interactions lead to biological insights into the disease.

scholarly journals Heterogeneity of Transcription Factor binding specificity models within and across cell lines

2015 ◽  
Author(s):  
Mahfuza Sharmin ◽  
Hector Corrada Bravo ◽  
Sridhar S. Hannenhalli
Keyword(s):  
Expression Patterns ◽  
Specific Interaction ◽  
Cell Types ◽  
Cell Type ◽  
Binding Interaction ◽  
Interaction Partners ◽  
A Cell ◽  
Cell Type Specific ◽  
Different Cell Types

Complex gene expression patterns are mediated by binding of transcription factors (TF) to specific genomic loci. The in vivo occupancy of a TF is, in large part, determined by the TFs DNA binding interaction partners, motivating genomic context based models of TF occupancy. However, the approaches thus far have assumed a uniform binding model to explain genome wide bound sites for a TF in a cell-type and as such heterogeneity of TF occupancy models, and the extent to which binding rules underlying a TFs occupancy are shared across cell types, has not been investigated. Here, we develop an ensemble based approach (TRISECT) to identify heterogeneous binding rules of cell-type specific TF occupancy and analyze the inter-cell-type sharing of such rules. Comprehensive analysis of 23 TFs, each with ChIP-Seq data in 4-12 cell-types, shows that by explicitly capturing the heterogeneity of binding rules, TRISECT accurately identifies in vivo TF occupancy (93%) substantially improving upon previous methods. Importantly, many of the binding rules derived from individual cell-types are shared across cell-types and reveal distinct yet functionally coherent putative target genes in different cell-types. Closer inspection of the predicted cell-type-specific interaction partners provides insights into context-specific functional landscape of a TF. Together, our novel ensemble-based approach reveals, for the first time, a widespread heterogeneity of binding rules, comprising interaction partners within a cell-type, many of which nevertheless transcend cell-types. Notably, the putative targets of shared binding rules in different cell-types, while distinct, exhibit significant functional coherence.

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