Jun Qiu1, Siqi Chen1, Lijuan Su1, Jinggong Liu1, Nannan Xiao1, Tian-Miao Ou1, Jia-Heng Tan1, Lian-Quan Gu2, Zhi-Shu Huang3, Ding Li4. 1. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou University City, 132 Waihuan East Road, Guangzhou 510006, PR China. 2. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou University City, 132 Waihuan East Road, Guangzhou 510006, PR China. Electronic address: cesglq@mail.sysu.edu.cn. 3. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou University City, 132 Waihuan East Road, Guangzhou 510006, PR China. Electronic address: ceshzs@mail.sysu.edu.cn. 4. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou University City, 132 Waihuan East Road, Guangzhou 510006, PR China. Electronic address: liding@mail.sysu.edu.cn.
Abstract
BACKGROUND: Cellular nucleic acid binding protein (CNBP) has been implicated in vertebrate craniofacial development and in myotonic dystrophy type 2 (DM2) and sporadic inclusion body myositis (sIBM) human diseases by controlling cell proliferation and survival to mediate neural crest expansion. CNBP has been found to bind single-stranded nucleic acid and promote rearrangements of nucleic acid secondary structure in an ATP-independent manner, acting as a nucleic acid chaperone. METHODS: A variety of methods were used, including cell viability assays, wound-scratch assays, chemotaxis assays, invasion assays, circular dichroic (CD) spectroscopy, NMR spectroscopy, chromatin immunoprecipitation, expression and purification of recombinant human CNBP, electrophoretic mobility shift assay (EMSA), surface plasmon resonance (SPR), fluorescence resonance energy transfer (FRET) analyses, luciferase reporter assay, Western blotting, and isothermal titration calorimetry (ITC). RESULTS: Up-regulation of CNBP induced human fibrosarcoma cell death and suppressed fibrosarcoma cell motility and invasiveness. It was found that CNBP transcriptionally down-regulated the expression of heterogeneous ribonucleoprotein K (hnRNP K) through its conversion of a G-rich sequence into G-quadruplex in the promoter of hnRNP K. G-quadruplex stabilizing ligand tetra-(N-methyl-4-pyridyl) porphyrin (TMPyP4) could interact with and stabilize the G-quadruplex, resulting in downregulation of hnRNP K transcription. CONCLUSIONS: CNBP overexpression caused increase of cell death and suppression of cell metastasis through its induction of G-quadruplex formation in the promoter of hnRNP K resulting in hnRNP K down-regulation. GENERAL SIGNIFICANCE: The present result provided a new solution for controlling hnRNP K expression, which should shed light on new anticancer drug design and development.
BACKGROUND:Cellular nucleic acid binding protein (CNBP) has been implicated in vertebrate craniofacial development and in myotonic dystrophy type 2 (DM2) and sporadic inclusion body myositis (sIBM) human diseases by controlling cell proliferation and survival to mediate neural crest expansion. CNBP has been found to bind single-stranded nucleic acid and promote rearrangements of nucleic acid secondary structure in an ATP-independent manner, acting as a nucleic acid chaperone. METHODS: A variety of methods were used, including cell viability assays, wound-scratch assays, chemotaxis assays, invasion assays, circular dichroic (CD) spectroscopy, NMR spectroscopy, chromatin immunoprecipitation, expression and purification of recombinant humanCNBP, electrophoretic mobility shift assay (EMSA), surface plasmon resonance (SPR), fluorescence resonance energy transfer (FRET) analyses, luciferase reporter assay, Western blotting, and isothermal titration calorimetry (ITC). RESULTS: Up-regulation of CNBP induced humanfibrosarcoma cell death and suppressed fibrosarcoma cell motility and invasiveness. It was found that CNBP transcriptionally down-regulated the expression of heterogeneous ribonucleoprotein K (hnRNP K) through its conversion of a G-rich sequence into G-quadruplex in the promoter of hnRNP K. G-quadruplex stabilizing ligand tetra-(N-methyl-4-pyridyl) porphyrin (TMPyP4) could interact with and stabilize the G-quadruplex, resulting in downregulation of hnRNP K transcription. CONCLUSIONS:CNBP overexpression caused increase of cell death and suppression of cell metastasis through its induction of G-quadruplex formation in the promoter of hnRNP K resulting in hnRNP K down-regulation. GENERAL SIGNIFICANCE: The present result provided a new solution for controlling hnRNP K expression, which should shed light on new anticancer drug design and development.
Authors: Aldana P David; Angélique Pipier; Federico Pascutti; Andrés Binolfi; Andrea M J Weiner; Emilse Challier; Sofía Heckel; Patrick Calsou; Dennis Gomez; Nora B Calcaterra; Pablo Armas Journal: Nucleic Acids Res Date: 2019-09-05 Impact factor: 16.971
Authors: Giacomo Canesin; Annalisa Di Ruscio; Mailin Li; Simone Ummarino; Andreas Hedblom; Reeham Choudhury; Agnieszka Krzyzanowska; Eva Csizmadia; Macarena Palominos; Anna Stiehm; Alexander Ebralidze; Shao-Yong Chen; Mahmoud A Bassal; Ping Zhao; Emanuela Tolosano; Laurence Hurley; Anders Bjartell; Daniel G Tenen; Barbara Wegiel Journal: Cell Rep Date: 2020-09-22 Impact factor: 9.423