A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.

Subtle alternative splicing leads to the formation of RNA variants lacking or including a small number of nucleotides. To date, the impact of subtle alternative splicing phenomena on protein biosynthesis has been studied in frame-preserving incidents. On the contrary, mRNA isoforms derived from fram...

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Main Authors: Evangelos D Karousis, Diamantis C Sideris
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2014-01-01
Series:PLoS ONE
Online Access:https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096557&type=printable
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author Evangelos D Karousis
Diamantis C Sideris
author_facet Evangelos D Karousis
Diamantis C Sideris
author_sort Evangelos D Karousis
collection DOAJ
description Subtle alternative splicing leads to the formation of RNA variants lacking or including a small number of nucleotides. To date, the impact of subtle alternative splicing phenomena on protein biosynthesis has been studied in frame-preserving incidents. On the contrary, mRNA isoforms derived from frame-shifting events were poorly studied and generally characterized as non-coding. This work provides evidence for a frame-shifting subtle alternative splicing event which results in the production of a novel protein isoform. We applied a combined molecular approach for the cloning and expression analysis of a human RNase κ transcript (RNase κ-02) which lacks four consecutive bases compared to the previously isolated RNase κ isoform. RNase κ-02 mRNA is expressed in all human cell lines tested end encodes the synthesis of a 134-amino-acid protein by utilizing an alternative initiation codon. The expression of RNase κ-02 in the cytoplasm of human cells was verified by Western blot and immunofluorescence analysis using a specific polyclonal antibody developed on the basis of the amino-acid sequence difference between the two protein isoforms. The results presented here show that subtle changes during mRNA splicing can lead to the expression of significantly altered protein isoforms.
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spelling doaj-art-ba9a04c1ce5a4e34b2782bb1352653c42025-08-20T02:14:42ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0195e9655710.1371/journal.pone.0096557A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.Evangelos D KarousisDiamantis C SiderisSubtle alternative splicing leads to the formation of RNA variants lacking or including a small number of nucleotides. To date, the impact of subtle alternative splicing phenomena on protein biosynthesis has been studied in frame-preserving incidents. On the contrary, mRNA isoforms derived from frame-shifting events were poorly studied and generally characterized as non-coding. This work provides evidence for a frame-shifting subtle alternative splicing event which results in the production of a novel protein isoform. We applied a combined molecular approach for the cloning and expression analysis of a human RNase κ transcript (RNase κ-02) which lacks four consecutive bases compared to the previously isolated RNase κ isoform. RNase κ-02 mRNA is expressed in all human cell lines tested end encodes the synthesis of a 134-amino-acid protein by utilizing an alternative initiation codon. The expression of RNase κ-02 in the cytoplasm of human cells was verified by Western blot and immunofluorescence analysis using a specific polyclonal antibody developed on the basis of the amino-acid sequence difference between the two protein isoforms. The results presented here show that subtle changes during mRNA splicing can lead to the expression of significantly altered protein isoforms.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096557&type=printable
spellingShingle Evangelos D Karousis
Diamantis C Sideris
A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.
PLoS ONE
title A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.
title_full A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.
title_fullStr A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.
title_full_unstemmed A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.
title_short A subtle alternative splicing event gives rise to a widely expressed human RNase k isoform.
title_sort subtle alternative splicing event gives rise to a widely expressed human rnase k isoform
url https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0096557&type=printable
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