| Texto completo | |
| Autor(es): |
Quel, Natalia G.
;
Rosa, Leonardo T.
;
Antonio, Larissa M.
;
Pinheiro, Glaucia M. S.
;
Barbosa, Leandro R. S.
;
Houry, Walid A.
;
Ramos, Carlos H. I.
Número total de Autores: 7
|
| Tipo de documento: | Artigo Científico |
| Fonte: | International Journal of Biological Macromolecules; v. 318, p. 11-pg., 2025-07-01. |
| Resumo | |
RUVBL1 and RUVBL2 proteins assemble into a heterohexameric ring and are essential for DNA repair in prokaryotes and chromatin homeostasis in eukaryotes. These proteins function as potential chaperones and ATPases. While most studies on eukaryotic RUVBL1/2 proteins have focused on human and yeast orthologs, they have revealed notable differences in conformational dynamics, protein interactions, and ATPase activity between these species. By investigating orthologs in other eukaryotic organisms, conserved features of RUVBL1/2 structure and function can be determined. In this study, we analyzed the genome of Aedes aegypti, a dipteran insect and a vector of Dengue, Zika, and Chikungunya viruses, to identify putative RUVBL1/2 family members. We identified protein sequences corresponding to RUVBL1 and RUVBL2, here named as AaRUVBL1 and AaRUVBL2. Purified recombinant AaRUVBL1/2 was properly folded and formed a hetero-dodecamer in solution. The complex exhibited enzymatic ATPase activity, confirming that these proteins are bona fide AAA+ ATPases. However, mutational analysis revealed that ATPase activity requires both AaRUVBL1 and AaRUVBL2, in contrast to the human ortholog, where RUVBL1 and RUVBL2 alone are active ATPases. To characterize the structure of the AaRUVBL1/2 complex, we combined SAXS and Cryo-EM techniques. Our findings indicate that the complex adopts a dodecameric barrel-shaped complex with a maximum dimension of similar to 16 nm. Single particle CryoEM analysis revealed a high degree of conformational heterogeneity, both between hexamer rings linked via the DII domains and among each hexameric ring. Overall, these findings contribute to a broader understanding of RUVBL1/2 proteins, particularly as this represents only the second structurally and functionally characterized RUVBL complex within the Animalia filum. (AU) | |
| Processo FAPESP: | 24/01429-5 - Investigação da relação estrutura-função das proteínas RPAP3, Pih1D1, WDR92 de Aedes aegypti e seu papel no desenvolvimento do inseto |
| Beneficiário: | Larissa Machado Antonio |
| Modalidade de apoio: | Bolsas no Brasil - Doutorado |
| Processo FAPESP: | 22/08855-4 - Estudo das interações entre componentes do chaperoma |
| Beneficiário: | Natália Galdi Quel |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |
| Processo FAPESP: | 22/11936-6 - Tripartite Tricarboxylate Transporters (TTT): investigando rede de transporte e sinalização no patógeno re-emergente Bordetella pertussis |
| Beneficiário: | Leonardo Talachia Rosa |
| Modalidade de apoio: | Auxílio à Pesquisa - Jovens Pesquisadores |
| Processo FAPESP: | 17/26131-5 - Chaperoma: estudo da relação entre a estrutura dos seus componentes e a manutenção da proteostase |
| Beneficiário: | Carlos Henrique Inacio Ramos |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |
| Processo FAPESP: | 18/11948-9 - Estudo do mecanismo de interação das co-chaperonas "J-domain proteins" com seus substratos e com a chaperona Hsp70 |
| Beneficiário: | Glaucia Melina Squizato Pinheiro de Castro |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |