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The transmission interface of the Saccharomyces cerevisiae multidrug transporter Pdr5: Val-656 located in intracellular loop 2 plays a major role in drug resistance.
Downes, Marianne T; Mehla, Jitender; Ananthaswamy, Neeti; Wakschlag, Adina; Lamonde, Micheala; Dine, Elliot; Ambudkar, Suresh V; Golin, John.
Afiliación
  • Downes MT; Department of Biology, The Catholic University of America, Washington, District of Columbia, USA.
Antimicrob Agents Chemother ; 57(2): 1025-34, 2013 Feb.
Article en En | MEDLINE | ID: mdl-23254431
Pdr5 is a major ATP-binding cassette (ABC) multidrug transporter regarded as the founding member of a fungal subfamily of clinically significant efflux pumps. When these proteins are overexpressed, they confer broad-spectrum ultraresistance. To better understand the evolution of these proteins under selective pressure, we exposed a Saccharomyces cerevisiae yeast strain already overexpressing Pdr5 to a lethal concentration of cycloheximide. This approach gave mutations that confer greater resistance to a subset of transport substrates. One of these mutations, V656L, is located in intracellular loop 2 (ICL2), a region predicted by structural studies with several other ABC transporters to play a critical role in the transmission interface between the ATP hydrolysis and drug transport domains. We show that this mutation increases drug resistance, possibly by altering the efficiency with which the energy from ATP hydrolysis is used for transport. Val-656 is a conserved residue, and an alanine substitution creates a nearly null phenotype for drug transport as well as reduced ATPase activity. We posit that despite its unusually small size, ICL2 is part of the transmission interface, and that alterations in this pathway can increase or decrease resistance to a broad spectrum of drugs.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Transporte de Membrana / Saccharomyces cerevisiae / Transportadoras de Casetes de Unión a ATP / Cicloheximida / Proteínas de Saccharomyces cerevisiae / Antifúngicos Idioma: En Revista: Antimicrob Agents Chemother Año: 2013 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas de Transporte de Membrana / Saccharomyces cerevisiae / Transportadoras de Casetes de Unión a ATP / Cicloheximida / Proteínas de Saccharomyces cerevisiae / Antifúngicos Idioma: En Revista: Antimicrob Agents Chemother Año: 2013 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos