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Translation of human tissue factor pathway inhibitor-? mRNA is controlled by alternative splicing within the 5' untranslated region.

ABSTRACT: Tissue factor pathway inhibitor (TFPI) blocks the initiation of coagulation by inhibiting TF-activated factor VII, activated factor X, and early prothrombinase. Humans produce two 3' splice variants, TFPI? and TFPI?, which are differentially expressed in endothelial cells and platelets and possess distinct structural features affecting their inhibitory function. TFPI also undergoes alternative splicing of exon 2 within its 5' untranslated region. The role of exon 2 splicing in translational regulation of human TFPI isoform expression is investigated.Exon 2 splicing occurs in TFPI? and TFPI? transcripts. Human tissue mRNA analysis uncovered a wide variability of exon 2 expression. Polysome analysis revealed a repressive effect of exon 2 on TFPI? translation but not on TFPI?. Luciferase reporter assays further exposed strong translational repression of TFPI? (90%) but not TFPI?. Use of a Morpholino to remove exon 2 from TFPI mRNA increased cell surface expression of endogenous TFPI?. Exon 2 also repressed luciferase production (80% to 90%) when paired with the ?-actin 3' untranslated region, suggesting that it is a general translational negative element whose effects are overcome by the TFPI? 3' untranslated region.Exon 2 is a molecular switch that prevents translation of TFPI?. This is the first demonstration of a 5' untranslated region alternative splicing event that alters translation of isoforms produced via independent 3' splicing events within the same gene. Therefore, it represents a previously unrecognized mechanism for translational control of protein expression. Differential expression of exon 2 denotes a mechanism to provide temporal and tissue-specific regulation of TFPI?-mediated anticoagulant activity.

SUBMITTER: Ellery PE

PROVIDER: S-EPMC4043743 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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Translation of human tissue factor pathway inhibitor-β mRNA is controlled by alternative splicing within the 5' untranslated region.

Ellery Paul E R PE Maroney Susan A SA Martinez Nicholas D ND Wickens Marvin P MP Mast Alan E AE

Arteriosclerosis, thrombosis, and vascular biology 20131114 1

<h4>Objective</h4>Tissue factor pathway inhibitor (TFPI) blocks the initiation of coagulation by inhibiting TF-activated factor VII, activated factor X, and early prothrombinase. Humans produce two 3' splice variants, TFPIα and TFPIβ, which are differentially expressed in endothelial cells and platelets and possess distinct structural features affecting their inhibitory function. TFPI also undergoes alternative splicing of exon 2 within its 5' untranslated region. The role of exon 2 splicing in ...[more]

PMID: 24233486

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Project description:Estrogens play an important role in breast cancer (BC) development and progression; when the two isoforms of the estrogen receptor (ER? and ER?) are co-expressed each of them mediate specific effects of these hormones in BC cells. ER? has been suggested to exert an antagonist role toward the oncogenic activities of ER?, and for this reason it is considered an oncosuppressor. As clinical evidence regarding a prognostic role for this receptor subtype in hormone-responsive BC is still limited and conflicting, more knowledge is required on the biological functions of ER? in cancer cells. We have previously described the ER? and ER? interactomes from BC cells, identifying specific and distinct patterns of protein interactions for the two receptors. In particular, we identified factors involved in mRNA splicing and maturation as important components of both ER? and ER? pathways. Guided by these findings, here we performed RNA sequencing to investigate in depth the differences in the early transcriptional events and RNA splicing patterns induced by estradiol in cells expressing ER? alone or ER? and ER?.Exon skipping was the most abundant splicing event in the post-transcriptional regulation by estradiol. We identified several splicing events induced by ER? alone and by ER?+ER?, demonstrating for the first time that ER? significantly affects estrogen-induced splicing in BC cells, as revealed by modification of a subset of ER?-dependent splicing by ER?, as well as by the presence of splicing isoforms only in ER?+cells. In particular, we observed that ER?+BC cell lines exhibited around 2-fold more splicing events than the ER?- cells. Interestingly, we identified putative direct targets of ER?-mediated alternative splicing by correlating the genomic locations of ER? and ER? binding sites with estradiol-induced differential splicing in the corresponding genes.Taken together, these results demonstrate that ER? significantly affects estrogen-induced early transcription and mRNA splicing in hormone-responsive BC cells, providing novel information on the biological role of ER? in these tumors.

Dataset Information

Translation of human tissue factor pathway inhibitor-? mRNA is controlled by alternative splicing within the 5' untranslated region.

Publications

Translation of human tissue factor pathway inhibitor-β mRNA is controlled by alternative splicing within the 5' untranslated region.

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