Leishmania major RUVBL1 has a hexameric conformation in solution and, in the presence of RUVBL2, forms a heterodimer with ATPase activity

Autor: Bárbara T. Amaro, Josielle Abrahão, Bárbara Ramalho Peres, Maria Isabel Nogueira Cano, Walid A. Houry, Annelize Zambon Barbosa Aragão, Carlos H.I. Ramos, Edna Gicela Ortiz Morea, Natália G. Quel
Přispěvatelé: Universidade Estadual de Campinas (UNICAMP), Universidade Estadual Paulista (Unesp), Univ Toronto
Rok vydání: 2021
Předmět:
Zdroj: Web of Science
Repositório Institucional da UNESP
Universidade Estadual Paulista (UNESP)
instacron:UNESP
ISSN: 1096-0384
Popis: Made available in DSpace on 2021-06-25T11:56:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-05-30 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) grant Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) CIHR 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. Univ Campinas UNICAMP, Inst Chem, BR-13083970 Campinas, SP, Brazil Sao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP, Brazil Univ Toronto, Dept Biochem, Toronto, ON M5G 1M1, Canada Univ Toronto, Dept Chem, Toronto, ON M5S 3H6, Canada Sao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP, Brazil FAPESP: 2012/50161-8 FAPESP: 2017/261315 FAPESP: 2013/10939-2 FAPESP: 2015/13521-4 FAPESP: 2014/25967-4 FAPESP: 2019/11496-3 CAPES: 99999.004913/2015-09 CIHR: PJT-173491
Databáze: OpenAIRE
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