Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei
The fate of an mRNA is determined by its interaction with proteins and small RNAs within dynamic complexes called ribonucleoprotein complexes (mRNPs). In Trypanosoma brucei and related kinetoplastids, responses to internal and external signals are mainly mediated by post-transcriptional processes. H...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
The Royal Society
2017-01-01
|
Series: | Open Biology |
Subjects: | |
Online Access: | https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.160159 |
id |
doaj-038a65615a434f23805c7029f765c113 |
---|---|
record_format |
Article |
spelling |
doaj-038a65615a434f23805c7029f765c1132020-11-25T03:28:26ZengThe Royal SocietyOpen Biology2046-24412017-01-017410.1098/rsob.160159160159Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma bruceiLuis Miguel De PablosSteve KellyJanaina de Freitas NascimentoJack SunterMark CarringtonThe fate of an mRNA is determined by its interaction with proteins and small RNAs within dynamic complexes called ribonucleoprotein complexes (mRNPs). In Trypanosoma brucei and related kinetoplastids, responses to internal and external signals are mainly mediated by post-transcriptional processes. Here, we used proximity-dependent biotin identification (BioID) combined with RNA-seq to investigate the changes resulting from ectopic expression of RBP10 and RBP9, two developmentally regulated RNA-binding proteins (RBPs). Both RBPs have reduced expression in insect procyclic forms (PCFs) compared with bloodstream forms (BSFs). Upon overexpression in PCFs, both proteins were recruited to cytoplasmic foci, co-localizing with the processing body marker SCD6. Further, both RBPs altered the transcriptome from a PCF- to a BSF-like pattern. Notably, upon expression of BirA*-RBP9 and BirA*-RBP10, BioID yielded more than 200 high confidence protein interactors (more than 10-fold enriched); 45 (RBP9) and 31 (RBP10) were directly related to mRNA metabolism. This study validates the use of BioID for investigating mRNP components but also illustrates the complexity of mRNP function.https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.160159rnagene expressiondifferentiationkinetoplastidbiotin identification |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Luis Miguel De Pablos Steve Kelly Janaina de Freitas Nascimento Jack Sunter Mark Carrington |
spellingShingle |
Luis Miguel De Pablos Steve Kelly Janaina de Freitas Nascimento Jack Sunter Mark Carrington Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei Open Biology rna gene expression differentiation kinetoplastid biotin identification |
author_facet |
Luis Miguel De Pablos Steve Kelly Janaina de Freitas Nascimento Jack Sunter Mark Carrington |
author_sort |
Luis Miguel De Pablos |
title |
Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei |
title_short |
Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei |
title_full |
Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei |
title_fullStr |
Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei |
title_full_unstemmed |
Characterization of RBP9 and RBP10, two developmentally regulated RNA-binding proteins in Trypanosoma brucei |
title_sort |
characterization of rbp9 and rbp10, two developmentally regulated rna-binding proteins in trypanosoma brucei |
publisher |
The Royal Society |
series |
Open Biology |
issn |
2046-2441 |
publishDate |
2017-01-01 |
description |
The fate of an mRNA is determined by its interaction with proteins and small RNAs within dynamic complexes called ribonucleoprotein complexes (mRNPs). In Trypanosoma brucei and related kinetoplastids, responses to internal and external signals are mainly mediated by post-transcriptional processes. Here, we used proximity-dependent biotin identification (BioID) combined with RNA-seq to investigate the changes resulting from ectopic expression of RBP10 and RBP9, two developmentally regulated RNA-binding proteins (RBPs). Both RBPs have reduced expression in insect procyclic forms (PCFs) compared with bloodstream forms (BSFs). Upon overexpression in PCFs, both proteins were recruited to cytoplasmic foci, co-localizing with the processing body marker SCD6. Further, both RBPs altered the transcriptome from a PCF- to a BSF-like pattern. Notably, upon expression of BirA*-RBP9 and BirA*-RBP10, BioID yielded more than 200 high confidence protein interactors (more than 10-fold enriched); 45 (RBP9) and 31 (RBP10) were directly related to mRNA metabolism. This study validates the use of BioID for investigating mRNP components but also illustrates the complexity of mRNP function. |
topic |
rna gene expression differentiation kinetoplastid biotin identification |
url |
https://royalsocietypublishing.org/doi/pdf/10.1098/rsob.160159 |
work_keys_str_mv |
AT luismigueldepablos characterizationofrbp9andrbp10twodevelopmentallyregulatedrnabindingproteinsintrypanosomabrucei AT stevekelly characterizationofrbp9andrbp10twodevelopmentallyregulatedrnabindingproteinsintrypanosomabrucei AT janainadefreitasnascimento characterizationofrbp9andrbp10twodevelopmentallyregulatedrnabindingproteinsintrypanosomabrucei AT jacksunter characterizationofrbp9andrbp10twodevelopmentallyregulatedrnabindingproteinsintrypanosomabrucei AT markcarrington characterizationofrbp9andrbp10twodevelopmentallyregulatedrnabindingproteinsintrypanosomabrucei |
_version_ |
1724584278308683776 |