scholarly journals Loss of PABPC1 is compensated by elevated PABPC4 and correlates with transcriptome changes

2021 ◽  
Author(s):  
Jingwei Xie ◽  
Xiaoyu Wei ◽  
Yu Chen
Keyword(s):  
Genome Editing ◽  
Transcriptome Analysis ◽  
Mrna Stability ◽  
Binding Protein ◽  
Transcriptional Regulators ◽  
Hek293 Cells ◽  
Translation Factor ◽  
Development And Differentiation ◽  
Insight Into

AbstractCytoplasmic poly(A) binding protein (PABP) is an essential translation factor that binds to the 3’ tail of mRNAs to promote translation and regulate mRNA stability. PABPC1 is the most abundant of several PABP isoforms that exist in mammals. Here, we used the CRISPR/Cas genome editing system to shift the isoform composition in HEK293 cells. Disruption of PABPC1 elevated PABPC4 levels. Transcriptome analysis revealed that the shift in the dominant PABP isoform was correlated with changes in key transcriptional regulators. This study provides insight into understanding the role of PABP isoforms in development and differentiation.

scholarly journals KH-type splicing regulatory protein is a new component of chromatoid body

Reproduction ◽  
2017 ◽  
Vol 154 (6) ◽  
pp. 723-733 ◽  
Author(s):  
Huijuan Zhang ◽  
Guishuan Wang ◽  
Lin Liu ◽  
Xiaolin Liang ◽  
Yu Lin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Germ Cell ◽  
Knockout Mice ◽  
Binding Protein ◽  
Regulatory Protein ◽  
Somatic Cells ◽  
Chromatoid Body ◽  
Mechanistic Insight ◽  
Insight Into

The chromatoid body (CB) is a specific cloud-like structure in the cytoplasm of haploid spermatids. Recent findings indicate that CB is identified as a male germ cell-specific RNA storage and processing center, but its function has remained elusive for decades. In somatic cells, KH-type splicing regulatory protein (KSRP) is involved in regulating gene expression and maturation of select microRNAs (miRNAs). However, the function of KSRP in spermatogenesis remains unclear. In this study, we showed that KSRP partly localizes in CB, as a component of CB. KSRP interacts with proteins (mouse VASA homolog (MVH), polyadenylate-binding protein 1 (PABP1) and polyadenylate-binding protein 2 (PABP2)), mRNAs (Tnp2 and Odf1) and microRNAs (microRNA-182) in mouse CB. Moreover, KSRP may regulate the integrity of CB via DDX5-miRNA-182 pathway. In addition, we found abnormal expressions of CB component in testes of Ksrp-knockout mice and of patients with hypospermatogenesis. Thus, our results provide mechanistic insight into the role of KSRP in spermatogenesis.

scholarly journals Regulation of thrombospondin-1 expression through AU-rich elements in the 3′UTR of the mRNA

2011 ◽  
Vol 16 (1) ◽  
Author(s):  
Asa Mcgray ◽  
Timothy Gingerich ◽  
James Petrik ◽  
Jonathan Lamarre
Keyword(s):  
Gene Expression ◽  
Mrna Stability ◽  
Half Life ◽  
Rna Binding ◽  
Binding Protein ◽  
Site Directed Mutagenesis ◽  
Hek293 Cells ◽  
Luciferase Reporter ◽  
Thrombospondin 1 ◽  
Mrna Half Life

AbstractThrombospondin-1 (TSP-1) is a matricellular protein that participates in numerous normal and pathological tissue processes and is rapidly modulated by different stimuli. The presence of 8 highly-conserved AU rich elements (AREs) within the 3′-untranslated region (3′UTR) of the TSP-1 mRNA suggests that post-transcriptional regulation is likely to represent one mechanism by which TSP-1 gene expression is regulated. We investigated the roles of these AREs, and proteins which bind to them, in the control of TSP-1 mRNA stability. The endogenous TSP-1 mRNA half-life is approximately 2.0 hours in HEK293 cells. Luciferase reporter mRNAs containing the TSP-1 3′UTR show a similar rate of decay, suggesting that the 3′UTR influences the decay rate. Site-directed mutagenesis of individual and adjacent AREs prolonged reporter mRNA halflife to between 2.2 and 4.4 hours. Mutation of all AREs increased mRNA half life to 8.8 hours, suggesting that all AREs have some effect, but that specific AREs may have key roles in stability regulation. A labeled RNA oligonucleotide derived from the most influential ARE was utilized to purify TSP-1 AREbinding proteins. The AU-binding protein AUF1 was shown to associate with this motif. These studies reveal that AREs in the 3′UTR control TSP-1 mRNA stability and that the RNA binding protein AUF1 participates in this control. These studies suggest that ARE-dependent control of TSP-1 mRNA stability may represent an important component in the control of TSP-1 gene expression.

Novel insight into the role of withering process in characteristic flavor formation of teas using transcriptome analysis and metabolite profiling

2019 ◽  
Vol 272 ◽  
pp. 313-322 ◽  
Author(s):  
Yu Wang ◽  
Peng-Cheng Zheng ◽  
Pan-Pan Liu ◽  
Xiao-Wei Song ◽  
Fei Guo ◽  
...  
Keyword(s):  
Transcriptome Analysis ◽  
Metabolite Profiling ◽  
Insight Into

scholarly journals Insight into the Role of Dynamics in the Conformational Switch of the Small GTP-binding Protein Arf1

2010 ◽  
Vol 285 (49) ◽  
pp. 37987-37994 ◽  
Author(s):  
Vanessa Buosi ◽  
Jean-Pierre Placial ◽  
Jean-Louis Leroy ◽  
Jacqueline Cherfils ◽  
Éric Guittet ◽  
...  
Keyword(s):  
Binding Protein ◽  
Gtp Binding Protein ◽  
Conformational Switch ◽  
Gtp Binding ◽  
Insight Into

scholarly journals Fructose modulates GLUT5 mRNA stability in differentiated Caco-2 cells: role of cAMP-signalling pathway and PABP (polyadenylated-binding protein)-interacting protein (Paip) 2

2003 ◽  
Vol 375 (1) ◽  
pp. 167-174 ◽  
Author(s):  
Florence GOUYON ◽  
Cercina ONESTO ◽  
Veronique DALET ◽  
Gilles PAGES ◽  
Armelle LETURQUE ◽  
...  
Keyword(s):  
Mrna Stability ◽  
Protein Complexes ◽  
Binding Protein ◽  
Interacting Protein ◽  
Camp Signalling ◽  
Camp Pathway ◽  
Post Transcriptional Regulation ◽  
Rna Protein Complexes ◽  
Camp Levels

In intestinal cells, levels of the fructose transporter GLUT5 are increased by glucose and to a greater extent by fructose. We investigated the mechanism by which fructose increases GLUT5 expression. In Caco-2 cells, fructose and glucose increased activity of the −2500/+41 GLUT5 promoter to the same extent. cAMP also activated the GLUT5 promoter. However, if a protein kinase A inhibitor was used to block cAMP signalling, extensive GLUT5 mRNA degradation was observed, with no change in basal transcription levels demonstrating the involvement of cAMP in GLUT5 mRNA stability. Indeed, the half-life of GLUT5 mRNA was correlated (R2=0.9913) with cellular cAMP levels. Fructose increased cAMP concentration more than glucose, accounting for the stronger effect of fructose when compared with that of glucose on GLUT5 production. We identified several complexes between GLUT5 3′-UTR RNA (where UTR stands for untranslated region) and cytosolic proteins that might participate in mRNA processing. Strong binding of a 140 kDa complex I was observed in sugar-deprived cells, with levels of binding lower in the presence of fructose and glucose by factors of 12 and 6 respectively. This may account for differences in the effects of fructose and glucose. In contrast, the amounts of two complexes of 96 and 48 kDa increased equally after stimulation with either glucose or fructose. Finally, PABP (polyadenylated-binding protein)-interacting protein 2, a destabilizing partner of PABP, was identified as a component of GLUT5 3′-UTR RNA–protein complexes. We conclude that the post-transcriptional regulation of GLUT5 by fructose involves increases in mRNA stability mediated by the cAMP pathway and Paip2 (PABP-interacting protein 2) binding.

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