An Exonic Switch Regulates Differential Accession of microRNAs to the Cd34 Transcript in Atherosclerosis Progression

Background: CD34+ Endothelial Progenitor Cells (EPCs) play an important role in the recovery of injured endothelium and contribute to atherosclerosis (ATH) pathogenesis. Previously we described a potential atherogenic role for miR-125 that we aimed to confirm in this work. Methods: Microarray hybridization, TaqMan Low Density Array (TLDA) cards, qPCR, and immunohistochemistry (IHC) were used to analyze expression of the miRNAs, proteins and transcripts here studied. Results: Here we have demonstrated an increase of resident CD34-positive cells in the aortic tissue of human and mice during ATH progression, as well as the presence of clusters of CD34-positive cells in the intima and adventitia of human ATH aortas. We introduce miR-351, which share the seed sequence with miR-125, as a potential effector of CD34. We show a splicing event at an internal/cryptic splice site at exon 8 of the murine Cd34 gene (exonic-switch) that would regulate the differential accession of miRNAs (including miR-125) to the coding region or to the 3’UTR of Cd34. Conclusions: We introduce new potential mediators of ATH progression (CD34 cell-clusters, miR-351), and propose a new mechanism of miRNA action, linked to a cryptic splicing site in the target-host gene, that would regulate the differential accession of miRNAs to their cognate binding sites.

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Multiple ASF/SF2 Sites in the Human Papillomavirus Type 16 (HPV-16) E4-Coding Region Promote Splicing to the Most Commonly Used 3′-Splice Site on the HPV-16 Genome

Journal of Virology ◽

10.1128/jvi.00462-10 ◽

2010 ◽

Vol 84 (16) ◽

pp. 8219-8230 ◽

Cited By ~ 33

Author(s):

Monika Somberg ◽

Stefan Schwartz

Keyword(s):

Cervical Cancer ◽

Human Papillomavirus ◽

Splice Site ◽

Binding Sites ◽

Mrna Levels ◽

Coding Region ◽

Cervical Lesions ◽

Hpv 16 ◽

Human Papillomavirus Type 16 ◽

E6 And E7

ABSTRACT Our results presented here demonstrate that the most abundant human papillomavirus type 16 (HPV-16) mRNAs expressing the viral oncogenes E6 and E7 are regulated by cellular ASF/SF2, itself defined as a proto-oncogene and overexpressed in cervical cancer cells. We show that the most frequently used 3′-splice site on the HPV-16 genome, site SA3358, which is used to produce primarily E4, E6, and E7 mRNAs, is regulated by ASF/SF2. Splice site SA3358 is immediately followed by 15 potential binding sites for the splicing factor ASF/SF2. Recombinant ASF/SF2 binds to the cluster of ASF/SF2 sites. Mutational inactivation of all 15 sites abolished splicing to SA3358 and redirected splicing to the downstream-located, late 3′-splice site SA5639. Overexpression of a mutant ASF/SF2 protein that lacks the RS domain, also totally inhibited the usage of SA3358 and redirected splicing to the late 3′-splice site SA5639. The 15 ASF/SF2 binding sites could be replaced by an ASF/SF2-dependent, HIV-1-derived splicing enhancer named GAR. This enhancer was also inhibited by the mutant ASF/SF2 protein that lacks the RS domain. Finally, silencer RNA (siRNA)-mediated knockdown of ASF/SF2 caused a reduction in spliced HPV-16 mRNA levels. Taken together, our results demonstrate that the major HPV-16 3′-splice site SA3358 is dependent on ASF/SF2. SA3358 is used by the most abundantly expressed HPV-16 mRNAs, including those encoding E6 and E7. High levels of ASF/SF2 may therefore be a requirement for progression to cervical cancer. This is supported by our earlier findings that ASF/SF2 is overexpressed in high-grade cervical lesions and cervical cancer.

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FAMILIAL GATA2 HAPLOINSUFFICIENCY CAUSED BY A NOVEL MUTATION INTRODUCING A CRYPTIC SPLICE SITE IN GATA2 EXON 3

10.26226/morressier.57bc1756d462b80290b4d80d ◽

2016 ◽

Author(s):

Claudia Wehr

Keyword(s):

Splice Site ◽

Novel Mutation ◽

Cryptic Splice Site

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Small dense low density lipoprotein has increased affinity for LDL receptor-independent cell surface binding sites: a potential mechanism for increased atherogenicity

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)32551-7 ◽

1998 ◽

Vol 39 (6) ◽

pp. 1263-1273 ◽

Cited By ~ 8

Author(s):

Narmer F. Galeano ◽

Maysoon Al-Haideri ◽

Fannie Keyserman ◽

Steven C. Rumsey ◽

Richard J. Deckelbaum

Keyword(s):

Cell Surface ◽

Binding Sites ◽

Low Density Lipoprotein ◽

Ldl Receptor ◽

Density Lipoprotein ◽

Low Density ◽

Potential Mechanism ◽

Surface Binding ◽

Cell Surface Binding

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Effects of cholestyramine on low density lipoprotein binding sites on liver membranes from rabbits with endogenous hypercholesterolemia induced by a wheat starch-casein diet.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)34825-7 ◽

1982 ◽

Vol 257 (7) ◽

pp. 3623-3627 ◽

Cited By ~ 7

Author(s):

Y Chao ◽

T T Yamin ◽

A W Alberts

Keyword(s):

Binding Sites ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Wheat Starch ◽

Low Density ◽

Casein Diet ◽

Liver Membranes ◽

Lipoprotein Binding

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Common variant of human NEDD4L activates a cryptic splice site to form a frameshifted transcript

Journal of Human Genetics ◽

10.1007/s100380200102 ◽

2002 ◽

Vol 47 (12) ◽

pp. 0665-0676 ◽

Cited By ~ 59

Author(s):

D. M. Dunn ◽

T. Ishigami ◽

J. Pankow ◽

A. von Niederhausern ◽

J. Alder ◽

...

Keyword(s):

Splice Site ◽

Common Variant ◽

Cryptic Splice Site

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Distinct Sets of Adjacent Heterogeneous Nuclear Ribonucleoprotein (hnRNP) A1/A2 Binding Sites Control 5′ Splice Site Selection in the hnRNP A1 mRNA Precursor

Journal of Biological Chemistry ◽

10.1074/jbc.m203633200 ◽

2002 ◽

Vol 277 (33) ◽

pp. 29745-29752 ◽

Cited By ~ 54

Author(s):

Stephen Hutchison ◽

Catherine LeBel ◽

Marco Blanchette ◽

Benoit Chabot

Keyword(s):

Splice Site ◽

Site Selection ◽

Binding Sites ◽

Hnrnp A1 ◽

Splice Site Selection ◽

Heterogeneous Nuclear Ribonucleoprotein ◽

Nuclear Ribonucleoprotein ◽

Mrna Precursor

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Mutational analysis of ectopic factor VIII transcripts from hemophilia A patients: identification of cryptic splice site, exon skipping and novel point mutations

Human Genetics ◽

10.1007/s004390050543 ◽

1997 ◽

Vol 100 (5-6) ◽

pp. 508-511 ◽

Cited By ~ 16

Author(s):

Kamiab Tavassoli ◽

Antonin Eigel ◽

Hartmut Pollmann ◽

Jürgen Horst

Keyword(s):

Factor Viii ◽

Splice Site ◽

Hemophilia A ◽

Mutational Analysis ◽

Point Mutations ◽

Exon Skipping ◽

Cryptic Splice Site

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Complete Thyroxine-Binding Globulin (TBG) Deficiency Produced by a Mutation in Acceptor Splice Site Causing Frameshift and Early Termination of Translation (TBG-Kankakee)12

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.83.10.5208 ◽

1998 ◽

Vol 83 (10) ◽

pp. 3604-3608

Author(s):

Gisah A. Carvalho ◽

Roy E. Weiss ◽

Samuel Refetoff

Keyword(s):

Splice Site ◽

Messenger Rna ◽

Restriction Site ◽

Splice Junction ◽

Early Termination ◽

Coding Region ◽

Acceptor Splice Site ◽

Exon 2 ◽

Gene Level ◽

Exon 1

Fourteen T4-binding globulin (TBG) variants have been identified at the gene level. They are all located in the coding region of the gene and 6 produce complete deficiency of TBG (TBG-CD). We now describe the first mutation in a noncoding region producing TBG-CD. The proband was treated for over 20 yr with L-T4 because of fatigue associated with a low concentration of serum total T4. Fifteen family members were studied showing low total T4 inherited as an X chromosome-linked trait, and affected males had undetectable TBG in serum. Sequencing of the entire coding region and promoter of the TBG gene revealed no abnormality. However, an A to G transition was found in the acceptor splice junction of intron II that produced a new HaeIII restriction site cosegregating with the TBG-CD phenotype. Sequencing exon 1 to exon 3 of TBG complementary DNA reverse transcribed from messenger RNA of skin fibroblasts from an affected male, confirmed a shift in the ag acceptor splice site. This results in the insertion of a G in exon 2 and causes a frameshift and a premature stop at codon 195. This early termination of translation predicts a truncated TBG lacking 201 amino acids.

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