Data/hora: 11/06/2024 08:36:41

Provedor de dados: | 74
País: | Brazil
Título: | Expression, purification and DNA-binding activities of two putative ModE proteins of Herbaspirillum seropedicae (Burkholderiales, Oxalobacteraceae)
Autores: | Souza,André L.F.; Chubatsu,Leda S.; Souza,Emanuel M.; Pedrosa,Fábio O.; Monteiro,Rose A.; Rego,Fabiane G.M.; Rigo,Liu U.
Data: | 2008-01-01
Ano: | 2008
Palavras-chave: | Herbaspirillum seropedicae; ModE1 protein; ModE2 protein.
Resumo: | In prokaryotes molybdenum is taken up by a high-affinity ABC-type transporter system encoded by the modABC genes. The endophyte β-Proteobacterium Herbaspirillum seropedicae has two modABC gene clusters and two genes encoding putative Mo-dependent regulator proteins (ModE1 and ModE2). Analysis of the amino acid sequence of the ModE1 protein of H. seropedicae revealed the presence of an N-terminal domain containing a DNA-binding helix-turn-helix motif (HTH) and a C-terminal domain with a molybdate-binding motif. The second putative regulator protein, ModE2, contains only the helix-turn-helix motif, similar to that observed in some sequenced genomes. We cloned the modE1 (810 bp) and modE2 (372 bp) genes and expressed them in Escherichia coli as His-tagged fusion proteins, which we subsequently purified. The over-expressed recombinant His-ModE1 was insoluble and was purified after solubilization with urea and then on-column refolded during affinity chromatography. The His-ModE2 was expressed as a soluble protein and purified by affinity chromatography. These purified proteins were analyzed by DNA band-shift assays using the modA2 promoter region as probe. Our results indicate that His-ModE1 and His-ModE2 are able to bind to the modA2 promoter region, suggesting that both proteins may play a role in the regulation of molybdenum uptake and metabolism in H. seropedicae.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572008000400022
Editor: | Sociedade Brasileira de Genética
Relação: | 10.1590/S1415-47572008000400022
Formato: | text/html
Fonte: | Genetics and Molecular Biology  v.31 n.3 2008
Fonte: | info:eu-repo/semantics/openAccess