Autor(es)

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]

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: Archives of Biochemistry and Biophysics; v. 703, MAY 30 2021.
• Citações Web of Science: 0

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. (AU)

• Processo FAPESP: 15/13521-4 - Caracterização funcional do complexo R2TP em diferentes organismos
• Beneficiário: Josielle Abrahão
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
• Processo FAPESP: 13/10939-2 - Caracterização de genes humanos como potencial chaperona com função desagregase
• Beneficiário: Josielle Abrahão
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 14/25967-4 - Caracterização do papel de domínios ATPase e das co-chaperonas Hsp40 e hop humanas nos mecanismos celulares de termotolerância e supressão de agregação proteica
• Beneficiário: Natália Galdi Quel
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 12/50161-8 - Estudo da estrutura e função da chaperona Hsp90 com ênfase no seu papel em homeostase celular
• Beneficiário: Carlos Henrique Inacio Ramos
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 19/11496-3 - PAPEL DO TERRA NA REGULAÇÃO DA MANUTENÇÃO DOS TELÔMEROS E NA SENESCÊNCIA REPLICATIVA EM Leishmania major
• Beneficiário: Edna Gicela Ortiz Morea
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado