Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.abb.2021.108841 http://hdl.handle.net/11449/209336 |
Resumo: | ATPases belonging to the AAA+ superfamily are associated with diverse cellular activities and are mainly characterized by a nucleotide-binding domain (NBD) containing the Walker A and Walker B motifs. AAA+ proteins have a range of functions, from DNA replication to protein degradation. Rvbs, also known as RUVBLs, are AAA+ ATPases with one NBD domain and were described from human to yeast as participants of the R2TP (Rvb1-Rvb2-Tah1-Pih1) complex. Although essential for the assembly of multiprotein complexes-containing DNA and RNA, the protozoa Rvb orthologs are less studied. For the first time, this work describes the Rvbs from Leishmania major, one of the causative agents of Tegumentar leishmaniasis in human. Recombinant LmRUVBL1 and LmRUVBL2 his-tagged proteins were successfully purified and investigated using biophysical tools. LmRUVBL1 was able to form a well-folded elongated hexamer in solution, while LmRUVBL2 formed a large aggregate. However, the co-expression of LmRUVBL1 and LmRUVBL2 assembled the proteins into an elongated heterodimer in solution. Thermo-stability and fluorescence experiments indicated that the LmRUVBL1/2 heterodimer had ATPase activity in vitro. This is an interesting result because hexameric LmRUVBL1 alone had low ATPase activity. Additionally, using independent SL-RNAseq libraries, it was possible to show that both proteins are expressed in all L. major life stages. Specific antibodies obtained against LmRUVBLs identified the proteins in promastigotes and metacyclics cell extracts. Together, the results here presented are the first step towards the characterization of Leishmania Rvbs, and may contribute to the development of possible strategies to intervene against leishmaniasis, a neglected tropical disease of great medical importance. |
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Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activityRvbLeishmania majorAAA plus proteinDNA helicasesATPaseATPases belonging to the AAA+ superfamily are associated with diverse cellular activities and are mainly characterized by a nucleotide-binding domain (NBD) containing the Walker A and Walker B motifs. AAA+ proteins have a range of functions, from DNA replication to protein degradation. Rvbs, also known as RUVBLs, are AAA+ ATPases with one NBD domain and were described from human to yeast as participants of the R2TP (Rvb1-Rvb2-Tah1-Pih1) complex. Although essential for the assembly of multiprotein complexes-containing DNA and RNA, the protozoa Rvb orthologs are less studied. For the first time, this work describes the Rvbs from Leishmania major, one of the causative agents of Tegumentar leishmaniasis in human. Recombinant LmRUVBL1 and LmRUVBL2 his-tagged proteins were successfully purified and investigated using biophysical tools. LmRUVBL1 was able to form a well-folded elongated hexamer in solution, while LmRUVBL2 formed a large aggregate. However, the co-expression of LmRUVBL1 and LmRUVBL2 assembled the proteins into an elongated heterodimer in solution. Thermo-stability and fluorescence experiments indicated that the LmRUVBL1/2 heterodimer had ATPase activity in vitro. This is an interesting result because hexameric LmRUVBL1 alone had low ATPase activity. Additionally, using independent SL-RNAseq libraries, it was possible to show that both proteins are expressed in all L. major life stages. Specific antibodies obtained against LmRUVBLs identified the proteins in promastigotes and metacyclics cell extracts. Together, the results here presented are the first step towards the characterization of Leishmania Rvbs, and may contribute to the development of possible strategies to intervene against leishmaniasis, a neglected tropical disease of great medical importance.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Foreign Affairs, Trade, and Development Canada (DFATD) grantConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)CIHRUniv Campinas UNICAMP, Inst Chem, BR-13083970 Campinas, SP, BrazilSao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP, BrazilUniv Toronto, Dept Biochem, Toronto, ON M5G 1M1, CanadaUniv Toronto, Dept Chem, Toronto, ON M5S 3H6, CanadaSao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP, BrazilFAPESP: 2012/50161-8FAPESP: 2017/261315FAPESP: 2013/10939-2FAPESP: 2015/13521-4FAPESP: 2014/25967-4FAPESP: 2019/11496-3CAPES: 99999.004913/2015-09CIHR: PJT-173491Elsevier B.V.Universidade Estadual de Campinas (UNICAMP)Universidade Estadual Paulista (Unesp)Univ TorontoAbrahao, JosielleAmaro, Barbara T.Peres, Barbara R.Quel, Natalia G.Araga, Annelize Z. B.Morea, Edna G. O. [UNESP]Cano, Maria Isabel N. [UNESP]Houry, Walid A.Ramos, Carlos H. I.2021-06-25T11:56:42Z2021-06-25T11:56:42Z2021-05-30info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article8http://dx.doi.org/10.1016/j.abb.2021.108841Archives Of Biochemistry And Biophysics. New York: Elsevier Science Inc, v. 703, 8 p., 2021.0003-9861http://hdl.handle.net/11449/20933610.1016/j.abb.2021.108841WOS:000641456700001Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengArchives Of Biochemistry And Biophysicsinfo:eu-repo/semantics/openAccess2021-10-23T19:28:03Zoai:repositorio.unesp.br:11449/209336Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T19:28:03Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
title |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
spellingShingle |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity Abrahao, Josielle Rvb Leishmania major AAA plus protein DNA helicases ATPase |
title_short |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
title_full |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
title_fullStr |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
title_full_unstemmed |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
title_sort |
Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity |
author |
Abrahao, Josielle |
author_facet |
Abrahao, Josielle Amaro, Barbara T. Peres, Barbara R. Quel, Natalia G. Araga, Annelize Z. B. Morea, Edna G. O. [UNESP] Cano, Maria Isabel N. [UNESP] Houry, Walid A. Ramos, Carlos H. I. |
author_role |
author |
author2 |
Amaro, Barbara T. Peres, Barbara R. Quel, Natalia G. Araga, Annelize Z. B. Morea, Edna G. O. [UNESP] Cano, Maria Isabel N. [UNESP] Houry, Walid A. Ramos, Carlos H. I. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual de Campinas (UNICAMP) Universidade Estadual Paulista (Unesp) Univ Toronto |
dc.contributor.author.fl_str_mv |
Abrahao, Josielle Amaro, Barbara T. Peres, Barbara R. Quel, Natalia G. Araga, Annelize Z. B. Morea, Edna G. O. [UNESP] Cano, Maria Isabel N. [UNESP] Houry, Walid A. Ramos, Carlos H. I. |
dc.subject.por.fl_str_mv |
Rvb Leishmania major AAA plus protein DNA helicases ATPase |
topic |
Rvb Leishmania major AAA plus protein DNA helicases ATPase |
description |
ATPases belonging to the AAA+ superfamily are associated with diverse cellular activities and are mainly characterized by a nucleotide-binding domain (NBD) containing the Walker A and Walker B motifs. AAA+ proteins have a range of functions, from DNA replication to protein degradation. Rvbs, also known as RUVBLs, are AAA+ ATPases with one NBD domain and were described from human to yeast as participants of the R2TP (Rvb1-Rvb2-Tah1-Pih1) complex. Although essential for the assembly of multiprotein complexes-containing DNA and RNA, the protozoa Rvb orthologs are less studied. For the first time, this work describes the Rvbs from Leishmania major, one of the causative agents of Tegumentar leishmaniasis in human. Recombinant LmRUVBL1 and LmRUVBL2 his-tagged proteins were successfully purified and investigated using biophysical tools. LmRUVBL1 was able to form a well-folded elongated hexamer in solution, while LmRUVBL2 formed a large aggregate. However, the co-expression of LmRUVBL1 and LmRUVBL2 assembled the proteins into an elongated heterodimer in solution. Thermo-stability and fluorescence experiments indicated that the LmRUVBL1/2 heterodimer had ATPase activity in vitro. This is an interesting result because hexameric LmRUVBL1 alone had low ATPase activity. Additionally, using independent SL-RNAseq libraries, it was possible to show that both proteins are expressed in all L. major life stages. Specific antibodies obtained against LmRUVBLs identified the proteins in promastigotes and metacyclics cell extracts. Together, the results here presented are the first step towards the characterization of Leishmania Rvbs, and may contribute to the development of possible strategies to intervene against leishmaniasis, a neglected tropical disease of great medical importance. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-06-25T11:56:42Z 2021-06-25T11:56:42Z 2021-05-30 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.abb.2021.108841 Archives Of Biochemistry And Biophysics. New York: Elsevier Science Inc, v. 703, 8 p., 2021. 0003-9861 http://hdl.handle.net/11449/209336 10.1016/j.abb.2021.108841 WOS:000641456700001 |
url |
http://dx.doi.org/10.1016/j.abb.2021.108841 http://hdl.handle.net/11449/209336 |
identifier_str_mv |
Archives Of Biochemistry And Biophysics. New York: Elsevier Science Inc, v. 703, 8 p., 2021. 0003-9861 10.1016/j.abb.2021.108841 WOS:000641456700001 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Archives Of Biochemistry And Biophysics |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
8 |
dc.publisher.none.fl_str_mv |
Elsevier B.V. |
publisher.none.fl_str_mv |
Elsevier B.V. |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1803650366825299968 |