Silence of Insulin-Like Growth Factor 2 mRNA-Binding Protein 1 Prevents Vascular Smooth Muscle Cells Proliferation via Nuclear Factor of Activated T Cells Isoform-3/Ca2+/Calmodulin Pathway

2021 ◽  
Vol 11 (4) ◽  
pp. 704-710
Author(s):  
Hu Tuo ◽  
Baozhen Yao ◽  
Bing He ◽  
Shiqian Yu ◽  
Danni Li ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Growth Factor ◽  
Nuclear Factor ◽  
Activated T Cells ◽  
Mrna Binding Protein ◽  
Intracellular Ca ◽  
Related Proteins ◽  
Mrna Binding ◽  
Vsmcs Proliferation

Increased proliferation of vascular smooth muscle cells (VSMCs) contributes to the pathogenesis of atherosclerosis (AS), and the insulin like growth factor 2 (IGF2) is involved in AS through effects on VSMCs growth and migration. The IGF2 mRNA-binding protein 1(IGF2BP1) is a secreted protein that can bind to IGF2 and regulate its localization, however, whether IGF2BP1 could regulate VSMCs proliferation remains to be elucidated. This study aimed to investigate the role of IGF2BP1 in VSMCs proliferation and uncover the potential mechanism. Primary human aortic VSMCs that transfected with or without shRNA-IGF2BP1 were stimulated by platelet-derived growth factor-BB (PDGF-BB), and then cell proliferation, intracellular Ca2+ level, cell apoptosis and the expression of IGF2BP1, calmodulin (CaM) and cell cycle-related proteins were detected. RNA pull down assay was used to determine the interaction between IGF2BP1 and nuclear factor of activated T cells isoform-3 (NFATc3). We found that PDGF-BB promoted cell proliferation and enhanced IGF2BP1 protein expression in a concentration-dependent manner. The 10 μg/L PDGF-BB significantly increased intracellular Ca2+ level, NFATc3, CaM and calcineurin A protein expression, TUNEL-positive cells, and the expression of cell cycle-related proteins cyclin D1/E1/B1. However, knockdown of IGF2BP1 significantly blunted all these effects induced by PDGF-BB. In addition, IGF2BP1 could bind to NFATc3 RNA. Collectively, knockdown of IGF2BP1 could inhibit PDGFBB- induced VSMCs proliferation via targeting NFATc3/Ca2+/calmodulin pathway and disturbing the effect of NFATc3/ on cell cycle.

scholarly journals Antisense-mediated loss of calcium homoeostasis endoplasmic reticulum protein (CHERP; ERPROT213-21) impairs Ca2+ mobilization, nuclear factor of activated T-cells (NFAT) activation and cell proliferation in Jurkat T-lymphocytes

2003 ◽  
Vol 373 (1) ◽  
pp. 133-143 ◽  
Author(s):  
Flavia A. O'ROURKE ◽  
Janice M. LaPLANTE ◽  
Maurice B. FEINSTEIN
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
T Cells ◽  
Endoplasmic Reticulum ◽  
T Lymphocytes ◽  
Nuclear Factor ◽  
Activated T Cells ◽  
Endoplasmic Reticulum Protein ◽  
Hel Cells ◽  
Er Membrane

We recently discovered a novel gene on chromosome 19p13.1 and its product, an integral endoplasmic reticulum (ER) membrane protein, termed CHERP (calcium homoeostasis endoplasmic reticulum protein). A monoclonal antibody against its C-terminal domain inhibits Ins(1,4,5)P3-induced Ca2+ release from ER membrane vesicles of many cell types, and an antisense-mediated knockdown of CHERP in human erythroleukemia (HEL) cells greatly impaired Ca2+ mobilization by thrombin. In the present paper, we explore further CHERP's function in Jurkat T-lymphocytes. Confocal laser immunofluorescence microscopy showed that CHERP was co-localized with the Ins(1,4,5)P3 receptor throughout the cytoplasmic and perinuclear region, as previously found in HEL cells. Transfection of Jurkat cells with a lacI-regulated mammalian expression vector containing CHERP antisense cDNA caused a knockdown of CHERP and impaired the rise of cytoplasmic Ca2+ (measured by fura-2 acetoxymethyl ester fluorescence) caused by phytohaemagglutinin (PHA) and thrombin. A 50% fall of CHERP decreased the PHA-induced rise of the cytoplasmic free Ca2+ concentration ([Ca2+]i), but Ca2+ influx was unaffected. Greater depletion of CHERP (>70%) did not affect the concentration of Ins(1,4,5)P3 receptors, but diminished the rise of [Ca2+]i in response to PHA to ≤30% of that in control cells, decreased Ca2+ influx and slowed the initial rate of [Ca2+]i rise caused by thapsigargin, an inhibitor of the sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase, suggesting there was also some deficit in ER Ca2+ stores. In CHERP-depleted cells the Ca2+-dependent activation and translocation of the key transcription factor NFAT (nuclear factor of activated T-cells) from cytoplasm to nucleus was suppressed. Furthermore, cell proliferation was greatly slowed (as in HEL cells) along with a 60% decrease in cyclin D1, a key regulator of progression through the G1 phase of the cell cycle. These findings provide further evidence that CHERP is an important component of the ER Ca2+-mobilizing system in cells, and its loss impairs Ca2+-dependent biochemical pathways and progression through the cell cycle.

Insulin-like growth factor 2 mRNA binding protein 2 promotes aerobic glycolysis and cell proliferation in pancreatic ductal adenocarcinoma via stabilizing GLUT1 mRNA

2019 ◽  
Vol 51 (7) ◽  
pp. 743-752 ◽  
Author(s):  
Shan Huang ◽  
Zheng Wu ◽  
Yunyun Cheng ◽  
Wenzhen Wei ◽  
Linlin Hao
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Pancreatic Ductal Adenocarcinoma ◽  
Binding Protein ◽  
Aerobic Glycolysis ◽  
Ductal Adenocarcinoma ◽  
Insulin Like Growth Factor ◽  
Mrna Binding Protein ◽  
Mrna Binding ◽  
Glut1 Mrna

Abstract Insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) is a member of the IGF2BP protein family consisting of IGF2BP1~3 with the capacity of binding to many transcripts and regulating RNA stability, localization, and translation. In this study, we discovered that expression of IGF2BP2 was upregulated and led to a poor prognosis in pancreatic ductal adenocarcinoma (PDAC). IGF2BP2 protein was gradually elevated from normal pancreas, pancreatic intraepithelial neoplasia to PDAC in an LSL-KrasG12D/+;LSL-Trp53R172H/+;Pdx1-Cre mouse model. Furthermore, we demonstrated that IGF2BP2 promoted aerobic glycolysis and PDAC cell proliferation through directly binding to and stabilizing GLUT1 mRNA. In summary, our study unveiled an important role of IGF2BP2 in PDAC development by modulating aerobic glycolysis and as a potential therapeutic target for PDAC treatment.

scholarly journals CDK1 couples proliferation with protein synthesis

2020 ◽  
Vol 219 (3) ◽  
Author(s):  
Katharina Haneke ◽  
Johanna Schott ◽  
Doris Lindner ◽  
Anne Kruse Hollensen ◽  
Christian Kroun Damgaard ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Protein Synthesis ◽  
Ribosomal Proteins ◽  
High Demand ◽  
Cell Synchronization ◽  
Translation Factors ◽  
Mrna Binding Protein ◽  
Mrna Binding ◽  
Global Translation

Cell proliferation exerts a high demand on protein synthesis, yet the mechanisms coupling the two processes are not fully understood. A kinase and phosphatase screen for activators of translation, based on the formation of stress granules in human cells, revealed cell cycle–associated kinases as major candidates. CDK1 was identified as a positive regulator of global translation, and cell synchronization experiments showed that this is an extramitotic function of CDK1. Different pathways including eIF2α, 4EBP, and S6K1 signaling contribute to controlling global translation downstream of CDK1. Moreover, Ribo-Seq analysis uncovered that CDK1 exerts a particularly strong effect on the translation of 5′TOP mRNAs, which includes mRNAs encoding ribosomal proteins and several translation factors. This effect requires the 5′TOP mRNA-binding protein LARP1, concurrent to our finding that LARP1 phosphorylation is strongly dependent on CDK1. Thus, CDK1 provides a direct means to couple cell proliferation with biosynthesis of the translation machinery and the rate of protein synthesis.

scholarly journals CDK1 couples proliferation with protein synthesis

2019 ◽  
Author(s):  
Katharina Haneke ◽  
Johanna Schott ◽  
Doris Lindner ◽  
Anne K. Hollensen ◽  
Christian K. Damgaard ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Protein Synthesis ◽  
Ribosomal Proteins ◽  
High Demand ◽  
Cell Synchronization ◽  
Translation Factors ◽  
Mrna Binding Protein ◽  
Mrna Binding ◽  
Global Translation

ABSTRACTCell proliferation exerts a high demand on protein synthesis, yet the mechanisms coupling the two processes are not fully understood. A kinase and phosphatase screen for activators of translation, based on the formation of stress granules in human cells, revealed cell cycle-associated kinases as major candidates. CDK1 was identified as a positive regulator of global translation, and cell synchronization experiments showed that this is an extra-mitotic function of CDK1. Dephosphorylation of eIF2α and S6K1 signaling were found to act downstream of CDK1. Moreover, Ribo-Seq analysis uncovered that CDK1 exerts a particularly strong effect on the translation of 5’TOP mRNAs, which includes mRNAs encoding for ribosomal proteins and several translation factors. This effect requires the 5’TOP mRNA-binding protein LARP1, concurrent to our finding that LARP1 phosphorylation is strongly dependent on CDK1. Taken together, our results show that CDK1 provides a direct means to couple cell proliferation with biosynthesis of the translation machinery and the rate of protein synthesis.

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