Cargando…
Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform
Endonuclease V (ENDOV) is a ribonuclease with affinity for inosine which is the deamination product of adenosine. The genomes of most organisms, including human, encode ENDOV homologs, yet knowledge about in vivo functions and gene regulation is sparse. To contribute in this field, we analyzed mRNA...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2019
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6839837/ https://www.ncbi.nlm.nih.gov/pubmed/31703097 http://dx.doi.org/10.1371/journal.pone.0225081 |
_version_ | 1783467496541519872 |
---|---|
author | Berges, Natalia Nawaz, Meh Sameen Børresdatter Dahl, Tuva Hagen, Lars Bjørås, Magnar Laerdahl, Jon K. Alseth, Ingrun |
author_facet | Berges, Natalia Nawaz, Meh Sameen Børresdatter Dahl, Tuva Hagen, Lars Bjørås, Magnar Laerdahl, Jon K. Alseth, Ingrun |
author_sort | Berges, Natalia |
collection | PubMed |
description | Endonuclease V (ENDOV) is a ribonuclease with affinity for inosine which is the deamination product of adenosine. The genomes of most organisms, including human, encode ENDOV homologs, yet knowledge about in vivo functions and gene regulation is sparse. To contribute in this field, we analyzed mRNA and protein expression of human ENDOV (hENDOV). Analyses of public sequence databases revealed numerous hENDOV transcript variants suggesting extensive alternative splicing. Many of the transcripts lacked one or more exons corresponding to conserved regions of the ENDOV core domain, suggesting that these transcripts do not encode for active proteins. Three complete transcripts were found with open reading frames encoding 282, 308 and 309 amino acids, respectively. Recombinant hENDOV 308 and hENDOV 309 share the same cleavage activity as hENDOV 282 which is the variant that has been used in previous studies of hENDOV. However, hENDOV 309 binds inosine-containing RNA with stronger affinity than the other isoforms. Overexpressed GFP-fused isoforms were found in cytoplasm, nucleoli and arsenite induced stress granules in human cells as previously reported for hENDOV 282. RT-qPCR analysis of the 3’-termini showed that hENDOV 308 and hENDOV 309 transcripts are more abundant than hENDOV 282 transcripts in immortalized cell lines, but not in primary cells, suggesting that cells regulate hENDOV mRNA expression. In spite of the presence of all three full-length transcripts, mass spectrometry analyses identified peptides corresponding to the hENDOV 309 isoform only. This result suggests that further studies of human ENDOV should rather encompass the hENDOV 309 isoform. |
format | Online Article Text |
id | pubmed-6839837 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-68398372019-11-15 Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform Berges, Natalia Nawaz, Meh Sameen Børresdatter Dahl, Tuva Hagen, Lars Bjørås, Magnar Laerdahl, Jon K. Alseth, Ingrun PLoS One Research Article Endonuclease V (ENDOV) is a ribonuclease with affinity for inosine which is the deamination product of adenosine. The genomes of most organisms, including human, encode ENDOV homologs, yet knowledge about in vivo functions and gene regulation is sparse. To contribute in this field, we analyzed mRNA and protein expression of human ENDOV (hENDOV). Analyses of public sequence databases revealed numerous hENDOV transcript variants suggesting extensive alternative splicing. Many of the transcripts lacked one or more exons corresponding to conserved regions of the ENDOV core domain, suggesting that these transcripts do not encode for active proteins. Three complete transcripts were found with open reading frames encoding 282, 308 and 309 amino acids, respectively. Recombinant hENDOV 308 and hENDOV 309 share the same cleavage activity as hENDOV 282 which is the variant that has been used in previous studies of hENDOV. However, hENDOV 309 binds inosine-containing RNA with stronger affinity than the other isoforms. Overexpressed GFP-fused isoforms were found in cytoplasm, nucleoli and arsenite induced stress granules in human cells as previously reported for hENDOV 282. RT-qPCR analysis of the 3’-termini showed that hENDOV 308 and hENDOV 309 transcripts are more abundant than hENDOV 282 transcripts in immortalized cell lines, but not in primary cells, suggesting that cells regulate hENDOV mRNA expression. In spite of the presence of all three full-length transcripts, mass spectrometry analyses identified peptides corresponding to the hENDOV 309 isoform only. This result suggests that further studies of human ENDOV should rather encompass the hENDOV 309 isoform. Public Library of Science 2019-11-08 /pmc/articles/PMC6839837/ /pubmed/31703097 http://dx.doi.org/10.1371/journal.pone.0225081 Text en © 2019 Berges et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
spellingShingle | Research Article Berges, Natalia Nawaz, Meh Sameen Børresdatter Dahl, Tuva Hagen, Lars Bjørås, Magnar Laerdahl, Jon K. Alseth, Ingrun Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform |
title | Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform |
title_full | Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform |
title_fullStr | Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform |
title_full_unstemmed | Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform |
title_short | Complex alternative splicing of human Endonuclease V mRNA, but evidence for only a single protein isoform |
title_sort | complex alternative splicing of human endonuclease v mrna, but evidence for only a single protein isoform |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6839837/ https://www.ncbi.nlm.nih.gov/pubmed/31703097 http://dx.doi.org/10.1371/journal.pone.0225081 |
work_keys_str_mv | AT bergesnatalia complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform AT nawazmehsameen complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform AT børresdatterdahltuva complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform AT hagenlars complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform AT bjørasmagnar complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform AT laerdahljonk complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform AT alsethingrun complexalternativesplicingofhumanendonucleasevmrnabutevidenceforonlyasingleproteinisoform |