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TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major

Leishmania major, a protozoan parasite that diverged early from the main eukaryotic lineage, exhibits unusual mechanisms of gene expression. Little is known in this organism about the transcription factors involved in the synthesis of tRNA, 5S rRNA, and snRNAs, transcribed by RNA Polymerase III (Pol...

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Autores principales: Román-Carraro, Fiordaliso C., Florencio-Martínez, Luis E., Romero-Meza, Gabriela, Nepomuceno-Mejía, Tomás, Carrero, Julio C., Arroyo, Rossana, Ortega-López, Jaime, Manning-Cela, Rebeca G., Martínez-Calvillo, Santiago
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6463643/
https://www.ncbi.nlm.nih.gov/pubmed/31058184
http://dx.doi.org/10.1155/2019/1425281
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author Román-Carraro, Fiordaliso C.
Florencio-Martínez, Luis E.
Romero-Meza, Gabriela
Nepomuceno-Mejía, Tomás
Carrero, Julio C.
Arroyo, Rossana
Ortega-López, Jaime
Manning-Cela, Rebeca G.
Martínez-Calvillo, Santiago
author_facet Román-Carraro, Fiordaliso C.
Florencio-Martínez, Luis E.
Romero-Meza, Gabriela
Nepomuceno-Mejía, Tomás
Carrero, Julio C.
Arroyo, Rossana
Ortega-López, Jaime
Manning-Cela, Rebeca G.
Martínez-Calvillo, Santiago
author_sort Román-Carraro, Fiordaliso C.
collection PubMed
description Leishmania major, a protozoan parasite that diverged early from the main eukaryotic lineage, exhibits unusual mechanisms of gene expression. Little is known in this organism about the transcription factors involved in the synthesis of tRNA, 5S rRNA, and snRNAs, transcribed by RNA Polymerase III (Pol III). Here we identify and characterize the TFIIIB subunit Bdp1 in L. major (LmBdp1). Bdp1 plays key roles in Pol III transcription initiation in other organisms, as it participates in Pol III recruitment and promoter opening. In silico analysis showed that LmBdp1 contains the typical extended SANT domain as well as other Bdp1 conserved regions. Nevertheless, LmBdp1 also displays distinctive features, including the presence of only one aromatic residue in the N-linker region. We were not able to produce null mutants of LmBdp1 by homologous recombination, as the obtained double replacement cell line contained an extra copy of LmBdp1, indicating that LmBdp1 is essential for the viability of L. major promastigotes. Notably, the mutant cell line showed reduced levels of the LmBdp1 protein, and its growth was significantly decreased in relation to wild-type cells. Nuclear run-on assays demonstrated that Pol III transcription was affected in the mutant cell line, and ChIP experiments showed that LmBdp1 binds to 5S rRNA, tRNA, and snRNA genes. Thus, our results indicate that LmBdp1 is an essential protein required for Pol III transcription in L. major.
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spelling pubmed-64636432019-05-05 TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major Román-Carraro, Fiordaliso C. Florencio-Martínez, Luis E. Romero-Meza, Gabriela Nepomuceno-Mejía, Tomás Carrero, Julio C. Arroyo, Rossana Ortega-López, Jaime Manning-Cela, Rebeca G. Martínez-Calvillo, Santiago Biomed Res Int Research Article Leishmania major, a protozoan parasite that diverged early from the main eukaryotic lineage, exhibits unusual mechanisms of gene expression. Little is known in this organism about the transcription factors involved in the synthesis of tRNA, 5S rRNA, and snRNAs, transcribed by RNA Polymerase III (Pol III). Here we identify and characterize the TFIIIB subunit Bdp1 in L. major (LmBdp1). Bdp1 plays key roles in Pol III transcription initiation in other organisms, as it participates in Pol III recruitment and promoter opening. In silico analysis showed that LmBdp1 contains the typical extended SANT domain as well as other Bdp1 conserved regions. Nevertheless, LmBdp1 also displays distinctive features, including the presence of only one aromatic residue in the N-linker region. We were not able to produce null mutants of LmBdp1 by homologous recombination, as the obtained double replacement cell line contained an extra copy of LmBdp1, indicating that LmBdp1 is essential for the viability of L. major promastigotes. Notably, the mutant cell line showed reduced levels of the LmBdp1 protein, and its growth was significantly decreased in relation to wild-type cells. Nuclear run-on assays demonstrated that Pol III transcription was affected in the mutant cell line, and ChIP experiments showed that LmBdp1 binds to 5S rRNA, tRNA, and snRNA genes. Thus, our results indicate that LmBdp1 is an essential protein required for Pol III transcription in L. major. Hindawi 2019-04-01 /pmc/articles/PMC6463643/ /pubmed/31058184 http://dx.doi.org/10.1155/2019/1425281 Text en Copyright © 2019 Fiordaliso C. Román-Carraro et al. https://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Román-Carraro, Fiordaliso C.
Florencio-Martínez, Luis E.
Romero-Meza, Gabriela
Nepomuceno-Mejía, Tomás
Carrero, Julio C.
Arroyo, Rossana
Ortega-López, Jaime
Manning-Cela, Rebeca G.
Martínez-Calvillo, Santiago
TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major
title TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major
title_full TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major
title_fullStr TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major
title_full_unstemmed TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major
title_short TFIIIB Subunit Bdp1 Participates in RNA Polymerase III Transcription in the Protozoan Parasite Leishmania major
title_sort tfiiib subunit bdp1 participates in rna polymerase iii transcription in the protozoan parasite leishmania major
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6463643/
https://www.ncbi.nlm.nih.gov/pubmed/31058184
http://dx.doi.org/10.1155/2019/1425281
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