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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

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

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Author(s):
Abrahao, Josielle [1] ; Amaro, Barbara T. [1] ; Peres, Barbara R. [1] ; Quel, Natalia G. [1] ; Araga, Annelize Z. B. [1] ; Morea, Edna G. O. [2] ; Cano, Maria Isabel N. [2] ; Houry, Walid A. [3, 4] ; Ramos, Carlos H. I. [1]
Total Authors: 9
Affiliation:
[1] Univ Campinas UNICAMP, Inst Chem, BR-13083970 Campinas, SP - Brazil
[2] Sao Paulo State Univ, Biosci Inst, Dept Chem & Biol Sci, BR-18618689 Botucatu, SP - Brazil
[3] Univ Toronto, Dept Chem, Toronto, ON M5S 3H6 - Canada
[4] Univ Toronto, Dept Biochem, Toronto, ON M5G 1M1 - Canada
Total Affiliations: 4
Document type: Journal article
Source: Archives of Biochemistry and Biophysics; v. 703, MAY 30 2021.
Web of Science Citations: 0
Abstract

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. (AU)

FAPESP's process: 15/13521-4 - Functional characterization of R2TP-Hsp90 complex in different organisms
Grantee:Josielle Abrahão
Support Opportunities: Scholarships abroad - Research Internship - Doctorate
FAPESP's process: 13/10939-2 - Characterization of human genes as potential chaperone with desagregase function
Grantee:Josielle Abrahão
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 14/25967-4 - Characterization of the role of ATPase domains and human co-chaperones Hsp40 and hop in cellular mechanisms of thermal tolerance and suppression of protein aggregation
Grantee:Natália Galdi Quel
Support Opportunities: Scholarships in Brazil - Doctorate
FAPESP's process: 12/50161-8 - Study of the structure and function of the Hsp90 chaperone with emphasis on its role in cellular homeostasis
Grantee:Carlos Henrique Inacio Ramos
Support Opportunities: Research Projects - Thematic Grants
FAPESP's process: 19/11496-3 - Role of TERRA in telomere regulation and replicative senescence in Leishmania major
Grantee:Edna Gicela Ortiz Morea
Support Opportunities: Scholarships in Brazil - Post-Doctoral