title: Dissecting biochemical peculiarities of the ATPase activity of TcSub2, a component of the mRNA export pathway in Trypanosoma cruzi.
abstract: The RNA helicase DEAD-box protein Sub2 (yeast)/UAP56 (mammals) is conserved across eukaryotes and is essential for mRNA export in trypanosomes. Despite the high conservation of Sub2 in lower eukaryotes such as Trypanosoma cruzi, the low conservation of other mRNA export factors raises questions regarding whether the mode of action of TcSub2 is similar to that of orthologs from other eukaryotes. Mutation of the conserved K87 residue of TcSub2 abolishes ATPase activity, showing that its ATPase domain is functional. However, the Vmax of TcSub2 was much higher than the Vmax described for the human protein UAP56, which suggests that the TcSub2 enzyme hydrolyzes ATP faster than its human homolog. Furthermore, we demonstrate that RNA association is less important to the activity of TcSub2 compared to UAP56. Our results show differences in activity of this protein, even though the structure of TcSub2 is very similar to UAP56. Functional complementation assays indicate that these differences may be common to other trypanosomatids. Distinct features of RNA influence and ATPase efficiency between UAP56 and TcSub2 may reflect distinct structures for functional sites of TcSub2. For this reason, ligand-based or structure-based methodologies can be applied to investigate the potential of TcSub2 as a target for new drugs.
authors.affiliation: Instituto Carlos Chagas, Fiocruz Paraná, Brazil.
authors.forename: Ize de Aguiar
authors.initials: IA
authors.lastname: Bittencourt
authors.affiliation: Instituto Carlos Chagas, Fiocruz Paraná, Brazil.
authors.forename: Mariana
authors.initials: M
authors.lastname: Serpeloni
authors.affiliation: Instituto Carlos Chagas, Fiocruz Paraná, Brazil.
authors.forename: Priscila Mazzochi
authors.initials: PM
authors.lastname: Hiraiwa
authors.affiliation: Instituto Carlos Chagas, Fiocruz Paraná, Brazil. Electronic address: tatianabrasil@fiocruz.br.
authors.forename: Tatiana
authors.initials: T
authors.lastname: de Arruda Campos Brasil de Souza
authors.affiliation: Instituto Carlos Chagas, Fiocruz Paraná, Brazil. Electronic address: aravila@fiocruz.br.
authors.forename: Andréa Rodrigues
authors.initials: AR
authors.lastname: Ávila
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D053487:DEAD-box RNA Helicases / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D009154:Mutation; D011487:Protein Conformation; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D013329:Structure-Activity Relationship; D014349:Trypanosoma cruzi / Q000201:enzymology*
chemical_list: D012333:RNA, Messenger; D000251:Adenosine Triphosphatases; C487886:DDX39B protein, human; C027900:RNA-dependent ATPase; D053487:DEAD-box RNA Helicases
keywords: ATPase; Trypanosoma cruzi; UAP56/Sub2