Deletion of the mouse GH-binding protein (mGHBP) mRNA polyadenylation and splicing sites does not abolish production of mGHBP

1997 ◽  
Vol 19 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Y Zhou ◽  
L He ◽  
G Baumann ◽  
J J Kopchick
Keyword(s):  
Genomic Sequence ◽  
Binding Protein ◽  
Mrna Splicing ◽  
Splice Acceptor Site ◽  
Acceptor Site ◽  
Carboxy Terminus ◽  
Regulation Of Expression ◽  
Gh Receptor ◽  
Selection Of

ABSTRACT In murine species, the GH receptor (mGHR) gene encodes a full-length membrane-anchored mGHR and a truncated soluble receptor ectodomain (the GH-binding protein; mGHBP). The mGHR and mGHBP mRNAs are generated by alternative pre-mRNA splicing. Similar GHR/GHBP pairs are generated in other species by proteolysis of the GHR. The regulation of GHBP expression and the biological role of GHBP are not clear. In order to begin to dissect the factors responsible for regulating expression of mGHR and mGHBP, we have cloned a mouse GH receptor/binding protein (mGHR/BP) minigene consisting of two mGHR cDNA fragments and an mGHR/BP genomic sequence, such that the mGHR and mGHBP can be derived from the minigene by mimicking native alternative pre-mRNA splicing. To study the possible role of selection of polyadenylation relative to the expression of mGHR and mGHBP, we deleted the two tandem poly A addition signals and the flanking AT-rich region within the exon (exon 8A) that encodes the carboxy terminus of mGHBP. In addition, two other mutated forms of the minigene, one containing a mutated alternative splice acceptor site (SAS) near exon 8A and the other possessing a deletion of the intron between exons 7 and 8A (intron 7/8A), were generated. Expression of the mGHR/BP minigene and its mutated forms in transfected mouse L cells revealed that removal of the polyadenylation signals diminished but did not abolish either mGHR or mGHBP production. However, mutation of the SAS yielded normal mGHR and an mGHBP which may be a result of the translation of an mRNA possessing an open reading frame in intron 7/8A. Additionally, removal of intron 7/8A abolished mGHR expression but resulted in mGHBP production. The results suggest that selection of alternative polyadenylation sites of the mGHR/BP gene does not play a major role in the regulation of expression of mGHR and mGHBP in vitro. These results also suggest that mutation of the SAS near exon 8A does not abolish the ability of mGHR/BP gene to produce an mGHBP that retains the ability to bind GH, although the new mGHBP may be different from the natural mGHBP at its carboxy terminus.

Association of CBFA2 mutation with decreased platelet PKC-θ and impaired receptor-mediated activation of GPIIb-IIIa and pleckstrin phosphorylation: proteins regulated by CBFA2 play a role in GPIIb-IIIa activation

Blood ◽  
2004 ◽  
Vol 103 (3) ◽  
pp. 948-954 ◽  
Author(s):  
Liansheng Sun ◽  
Guangfen Mao ◽  
A. Koneti Rao
Keyword(s):  
Molecular Mechanisms ◽  
Splice Acceptor Site ◽  
Heterozygous Mutation ◽  
Acceptor Site ◽  
Runt Domain ◽  
Platelet Protein ◽  
Binding Factor ◽  
Reported Data ◽  
And Function

AbstractThe mechanisms by which agonists activate glycoprotein (GP) IIb-IIIa function remain unclear. We have reported data on a patient with thrombocytopenia and impaired receptor-mediated aggregation, phosphorylation of pleckstrin (a protein kinase C [PKC] substrate), and activation of the GPIIb-IIIa complex. Abnormalities in hematopoietic transcription factors have been associated with thrombocytopenia and platelet dysfunction. To define the molecular mechanisms, we amplified from patient platelet RNA exons 3 to 6 of core-binding factor A2 (CBFA2) cDNA, which encompasses the DNA-binding Runt domain; a 13-nucleotide (nt) deletion was found (796-808 nt). The gDNA revealed a heterozygous mutation (G>T) in intron 3 at the splice acceptor site for exon 4, leading to a frameshift with premature termination in the Runt domain. On immunoblotting, platelet CBFA2, PKC-θ, albumin, and IgG were decreased, but pleckstrin, PKC-α, -βI, -βII, -η, -ϵ, -δ, and -ζ, and fibrinogen were normal. Our conclusions are that (1) CBFA2 mutation is associated with not only thrombocytopenia, but also impaired platelet protein phosphorylation and GPIIb-IIIa activation; (2) proteins regulated by CBFA2 are required for inside-out signal transduction-dependent activation of GPIIb-IIIa; and (3) we have documented the first deficiency of a human PKC isozyme (PKC-θ), suggesting a major role of this isozyme in platelet production and function. (Blood. 2004;103:948-954)

Potential therapeutic strategies for targeting Y-box-binding protein 1 in cancers

2021 ◽  
Vol 21 ◽  
Author(s):  
Jia-Wei Yang ◽  
Chao Sun ◽  
Qiu-Yang Jin ◽  
Xing-Hui Qiao ◽  
Xiu-Li Guo
Keyword(s):  
Nuclear Translocation ◽  
Binding Protein ◽  
Circular Rna ◽  
Therapeutic Strategies ◽  
Mrna Splicing ◽  
Tumor Proliferation ◽  
Normal Cells ◽  
The Past ◽  
Potential Cancer

: As one of the most conservative proteins in evolution, Y-box-binding protein 1 (YB-1) has long been considered as a potential cancer target. YB-1 is usually poorly expressed in normal cells and exerts cellular physiological functions such as DNA repair, pre-mRNA splicing and mRNA stabilizing. In cancer cells, the expression of YB-1 is up-regulated and undergoes nuclear translocation and contributes to tumorigenesis, angiogenesis, tumor proliferation, invasion, migration and chemotherapy drug resistance. During the past decades, a variety of pharmacological tools such as siRNA, shRNA, microRNA, circular RNA, lncRNA and various compounds have been developed to target YB-1 for cancer therapy. In this review, we describe the physiological characteristics of YB-1 in detail, highlight the role of YB-1 in tumors and summarize the current therapeutic methods for targeting YB-1 in cancer.

scholarly journals Corrigendum: A Role of U12 Intron in Proper Pre-mRNA Splicing of Plant Cap Binding Protein 20 Genes

2019 ◽  
Vol 10 ◽  
Author(s):  
Marcin Pieczynski ◽  
Katarzyna Kruszka ◽  
Dawid Bielewicz ◽  
Jakub Dolata ◽  
Michal Szczesniak ◽  
...  
Keyword(s):  
Binding Protein ◽  
Mrna Splicing

Enigmatic rhodopsin mutation creates an exceptionally strong splice acceptor site

2019 ◽  
Vol 29 (2) ◽  
pp. 295-304
Author(s):  
Lisa M Riedmayr ◽  
Sybille Böhm ◽  
Martin Biel ◽  
Elvir Becirovic
Keyword(s):  
Branch Point ◽  
Protein Level ◽  
Mrna Splicing ◽  
Hek293 Cells ◽  
Splice Acceptor Site ◽  
Mouse Retina ◽  
Acceptor Site ◽  
Splice Acceptor ◽  
The Core ◽  
The Impact

Abstract The c.620 T > G mutation in rhodopsin found in the first mapped autosomal dominant retinitis pigmentosa (adRP) locus is associated with severe, early-onset RP. Intriguingly, another mutation affecting the same nucleotide (c.620 T > A) is related to a mild, late-onset RP. Assuming that both mutations are missense mutations (Met207Arg and Met207Lys) hampering the ligand-binding pocket, previous work addressed how they might differentially impair rhodopsin function. Here, we investigated the impact of both mutations at the mRNA and protein level in HEK293 cells and in the mouse retina. We show that, in contrast to c.620 T > A, c.620 T > G is a splicing mutation, which generates an exceptionally strong splice acceptor site (SAS) resulting in a 90 bp in-frame deletion and protein mislocalization in vitro and in vivo. Moreover, we identified the core element underlying the c.620 T > G SAS strength. Finally, we demonstrate that the c.620 T > G SAS is very flexible in branch point choice, which might explain its remarkable performance. Based on these results, we suggest that (i) point mutations should be routinely tested for mRNA splicing to avoid dispensable analysis of mutations on protein level, which do not naturally exist. (ii) Puzzling disease courses of mutations in other genes might also correlate with their effects on mRNA splicing. (iii) Flexibility in branch point choice might be another factor influencing the SAS strength. (iv) The core splice element identified in this study could be useful for biotechnological applications requiring effective SAS.

scholarly journals Pbp1p, a Factor Interacting withSaccharomyces cerevisiae Poly(A)-Binding Protein, Regulates Polyadenylation

1998 ◽  
Vol 18 (12) ◽  
pp. 7383-7396 ◽  
Author(s):  
David A. Mangus ◽  
Nadia Amrani ◽  
Allan Jacobson
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Functional Role ◽  
Binding Protein ◽  
Carboxy Terminus ◽  
Ribosomal Subunits ◽  
Initiation Of Translation ◽  
Genes Encoding ◽  
Key Factor ◽  
Two Hybrid

ABSTRACT The poly(A) tail of an mRNA is believed to influence the initiation of translation, and the rate at which the poly(A) tail is removed is thought to determine how fast an mRNA is degraded. One key factor associated with this 3′-end structure is the poly(A)-binding protein (Pab1p) encoded by the PAB1 gene inSaccharomyces cerevisiae. In an effort to learn more about the functional role of this protein, we used a two-hybrid screen to determine the factor(s) with which it interacts. We identified five genes encoding factors that specifically interact with the carboxy terminus of Pab1p. Of a total of 44 specific clones identified,PBP1 (for Pab1p-binding protein) was isolated 38 times. Of the putative interacting genes examined, PBP1 promoted the highest level of resistance to 3-aminotriazole (>100 mM) in constructs in which HIS3 was used as a reporter. We determined that a fraction of Pbp1p cosediments with polysomes in sucrose gradients and that its distribution is very similar to that of Pab1p. Disruption ofPBP1 showed that it is not essential for viability but can suppress the lethality associated with a PAB1 deletion. The suppression of pab1Δ by pbp1Δ appears to be different from that mediated by other pab1 suppressors, since disruption of PBP1 does not alter translation rates, affect accumulation of ribosomal subunits, change mRNA poly(A) tail lengths, or result in a defect in mRNA decay. Rather, Pbp1p appears to function in the nucleus to promote proper polyadenylation. In the absence of Pbp1p, 3′ termini of pre-mRNAs are properly cleaved but lack full-length poly(A) tails. These effects suggest that Pbp1p may act to repress the ability of Pab1p to negatively regulate polyadenylation.

Hormonal regulation of expression of ileal bile acid binding protein in suckling rats

2000 ◽  
Vol 278 (6) ◽  
pp. R1555-R1563 ◽  
Author(s):  
Sandy T. Hwang ◽  
Susan J. Henning
Keyword(s):  
Bile Acid ◽  
Hormonal Regulation ◽  
Normal Development ◽  
Binding Protein ◽  
Secondary Response ◽  
Regulation Of Expression ◽  
Acid Binding Protein ◽  
Bile Acid Binding ◽  
Developing Rat

Ileal bile acid binding protein (IBABP) is a cytosolic protein believed to be involved in the absorption of conjugated bile acids. In rodents this protein and its mRNA have been shown to increase markedly during the third postnatal week. Because this period of ontogeny is characterized by increasing circulating concentrations of glucocorticoids and thyroxine, the goal of our study was to investigate the role of these hormones in IBABP expression in the developing rat. Administration of various doses of dexamethasone (Dex) during the second postnatal week caused a robust induction of IBABP mRNA and protein. Plateau levels of IBABP mRNA occurred at a Dex dose of 0.1 μg/g body wt, which is within the physiological range. IBABP mRNA was not appreciably induced until 24 h after treatment, suggesting that glucocorticoids influence IBABP either through a delayed primary or a secondary response mechanism. The regional pattern of IBABP mRNA elicited by Dex mimicked that seen during normal development, with appearance in distal ileum preceding proximal ileum. Thyroxine injections did not result in a significant increase of IBABP mRNA, and synergism between Dex and thyroxine was not observed. Taken together, our data suggest that maturation of IBABP expression is influenced by glucocorticoids but not by thyroxine.

scholarly journals First person – Andria A. Lytridou

2020 ◽  
Vol 133 (20) ◽  
pp. jcs255208
Keyword(s):  
Endoplasmic Reticulum ◽  
Endoplasmic Reticulum Stress ◽  
Binding Protein ◽  
Early Career ◽  
First Person ◽  
Early Career Researchers ◽  
Selection Of

ABSTRACTFirst Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping early-career researchers promote themselves alongside their papers. Andria A. Lytridou is first author on ‘Stbd1 promotes glycogen clustering during endoplasmic reticulum stress and supports survival of mouse myoblasts’, published in JCS. Andria is a PhD student in the lab of Dr Petros P. Petrou at The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus, investigating the role of the glycogen-binding protein Stbd1 in physiological and pathological processes.

scholarly journals A Role of U12 Intron in Proper Pre-mRNA Splicing of Plant Cap Binding Protein 20 Genes

2018 ◽  
Vol 9 ◽  
Author(s):  
Marcin Pieczynski ◽  
Katarzyna Kruszka ◽  
Dawid Bielewicz ◽  
Jakub Dolata ◽  
Michal Szczesniak ◽  
...  
Keyword(s):  
Binding Protein ◽  
Mrna Splicing
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