Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins.
Poly(A) Binding Proteins (PABPs) are major eukaryotic RNA-binding proteins (RBPs) with multiple roles associated with mRNA stability and translation and characterized mainly from multicellular organisms and yeasts. A variable number of PABP homologues are seen in different organisms however the biol...
Published in: | PLOS Neglected Tropical Diseases |
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2021
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Online Access: | https://doi.org/10.1371/journal.pntd.0009899 https://doaj.org/article/7582158e93974ec7b0804d1be9beaee2 |
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ftdoajarticles:oai:doaj.org/article:7582158e93974ec7b0804d1be9beaee2 2023-05-15T15:12:20+02:00 Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. Ludmila A Assis Moezio V C Santos Filho Joao R da Cruz Silva Maria J R Bezerra Irassandra R P U C de Aquino Kleison C Merlo Fabiola B Holetz Christian M Probst Antonio M Rezende Barbara Papadopoulou Tamara D C da Costa Lima Osvaldo P de Melo Neto 2021-10-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0009899 https://doaj.org/article/7582158e93974ec7b0804d1be9beaee2 EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0009899 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009899 https://doaj.org/article/7582158e93974ec7b0804d1be9beaee2 PLoS Neglected Tropical Diseases, Vol 15, Iss 10, p e0009899 (2021) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2021 ftdoajarticles https://doi.org/10.1371/journal.pntd.0009899 2022-12-31T11:07:09Z Poly(A) Binding Proteins (PABPs) are major eukaryotic RNA-binding proteins (RBPs) with multiple roles associated with mRNA stability and translation and characterized mainly from multicellular organisms and yeasts. A variable number of PABP homologues are seen in different organisms however the biological reasons for multiple PABPs are generally not well understood. In the unicellular Leishmania, dependent on post-transcriptional mechanisms for the control of its gene expression, three distinct PABPs are found, with yet undefined functional distinctions. Here, using RNA-immunoprecipitation sequencing analysis we show that the Leishmania PABP1 preferentially associates with mRNAs encoding ribosomal proteins, while PABP2 and PABP3 bind to an overlapping set of mRNAs distinct to those enriched in PABP1. Immunoprecipitation studies combined to mass-spectrometry analysis identified RBPs differentially associated with PABP1 or PABP2, including RBP23 and DRBD2, respectively, that were investigated further. Both RBP23 and DRBD2 bind directly to the three PABPs in vitro, but reciprocal experiments confirmed preferential co-immunoprecipitation of PABP1, as well as the EIF4E4/EIF4G3 based translation initiation complex, with RBP23. Other RBP23 binding partners also imply a direct role in translation. DRBD2, in contrast, co-immunoprecipitated with PABP2, PABP3 and with RBPs unrelated to translation. Over 90% of the RBP23-bound mRNAs code for ribosomal proteins, mainly absent from the transcripts co-precipitated with DRBD2. These experiments suggest a novel and specific route for translation of the ribosomal protein mRNAs, mediated by RBP23, PABP1 and the associated EIF4E4/EIF4G3 complex. They also highlight the unique roles that different PABP homologues may have in eukaryotic cells associated with mRNA translation. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 15 10 e0009899 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Ludmila A Assis Moezio V C Santos Filho Joao R da Cruz Silva Maria J R Bezerra Irassandra R P U C de Aquino Kleison C Merlo Fabiola B Holetz Christian M Probst Antonio M Rezende Barbara Papadopoulou Tamara D C da Costa Lima Osvaldo P de Melo Neto Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Poly(A) Binding Proteins (PABPs) are major eukaryotic RNA-binding proteins (RBPs) with multiple roles associated with mRNA stability and translation and characterized mainly from multicellular organisms and yeasts. A variable number of PABP homologues are seen in different organisms however the biological reasons for multiple PABPs are generally not well understood. In the unicellular Leishmania, dependent on post-transcriptional mechanisms for the control of its gene expression, three distinct PABPs are found, with yet undefined functional distinctions. Here, using RNA-immunoprecipitation sequencing analysis we show that the Leishmania PABP1 preferentially associates with mRNAs encoding ribosomal proteins, while PABP2 and PABP3 bind to an overlapping set of mRNAs distinct to those enriched in PABP1. Immunoprecipitation studies combined to mass-spectrometry analysis identified RBPs differentially associated with PABP1 or PABP2, including RBP23 and DRBD2, respectively, that were investigated further. Both RBP23 and DRBD2 bind directly to the three PABPs in vitro, but reciprocal experiments confirmed preferential co-immunoprecipitation of PABP1, as well as the EIF4E4/EIF4G3 based translation initiation complex, with RBP23. Other RBP23 binding partners also imply a direct role in translation. DRBD2, in contrast, co-immunoprecipitated with PABP2, PABP3 and with RBPs unrelated to translation. Over 90% of the RBP23-bound mRNAs code for ribosomal proteins, mainly absent from the transcripts co-precipitated with DRBD2. These experiments suggest a novel and specific route for translation of the ribosomal protein mRNAs, mediated by RBP23, PABP1 and the associated EIF4E4/EIF4G3 complex. They also highlight the unique roles that different PABP homologues may have in eukaryotic cells associated with mRNA translation. |
format |
Article in Journal/Newspaper |
author |
Ludmila A Assis Moezio V C Santos Filho Joao R da Cruz Silva Maria J R Bezerra Irassandra R P U C de Aquino Kleison C Merlo Fabiola B Holetz Christian M Probst Antonio M Rezende Barbara Papadopoulou Tamara D C da Costa Lima Osvaldo P de Melo Neto |
author_facet |
Ludmila A Assis Moezio V C Santos Filho Joao R da Cruz Silva Maria J R Bezerra Irassandra R P U C de Aquino Kleison C Merlo Fabiola B Holetz Christian M Probst Antonio M Rezende Barbara Papadopoulou Tamara D C da Costa Lima Osvaldo P de Melo Neto |
author_sort |
Ludmila A Assis |
title |
Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. |
title_short |
Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. |
title_full |
Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. |
title_fullStr |
Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. |
title_full_unstemmed |
Identification of novel proteins and mRNAs differentially bound to the Leishmania Poly(A) Binding Proteins reveals a direct association between PABP1, the RNA-binding protein RBP23 and mRNAs encoding ribosomal proteins. |
title_sort |
identification of novel proteins and mrnas differentially bound to the leishmania poly(a) binding proteins reveals a direct association between pabp1, the rna-binding protein rbp23 and mrnas encoding ribosomal proteins. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2021 |
url |
https://doi.org/10.1371/journal.pntd.0009899 https://doaj.org/article/7582158e93974ec7b0804d1be9beaee2 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 15, Iss 10, p e0009899 (2021) |
op_relation |
https://doi.org/10.1371/journal.pntd.0009899 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009899 https://doaj.org/article/7582158e93974ec7b0804d1be9beaee2 |
op_doi |
https://doi.org/10.1371/journal.pntd.0009899 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
15 |
container_issue |
10 |
container_start_page |
e0009899 |
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1766343032894914560 |