Acanthamoeba alternative oxidase genes: Identification, characterisation and potential as antimicrobial targets

Fiona Henriquez-Mui, James McBride, Sara J. Campbell, Tania Ramos, Paul R. Ingram, Fiona Roberts, Sinead Tinney, Craig W. Roberts

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Alternative oxidase (AOX) is a mitochondrial protein that acts as an alternative terminal oxidase to the conventional cytochrome oxidases. It is present in certain prokaryotes, plants, fungi and some protozoa but absent in mammals. AOX activity has previously been described in Acanthamoeba, although no genetic evidence has been reported. Herein, two AOX (AcAOX) genes designated isoforms A and B, were obtained from Acanthamoeba castellanii by a combination of degenerate PCR from cDNA and a series of 51 and 3' rapid amplification of cDNA ends. The corresponding genomic sequences of these AcAOXs were also obtained. Each gene spans six exons over a region of 1607 and 1619 bp, respectively. Isoforms A and B have open reading frames of 1113 and 1125 bp, respectively. Each encodes a protein with a predicted molecular weight of 42 kDa. Each AcAOX protein has a predicted cleavable mitochondrial targeting sequence. The full-length AcAOX is functionally active as it complements hemL-deficient Escherichia coli and inhibited by the inhibitor of ACX, salicylhydroxamic acid (SHAM). SHAM is effective against A. castellanii and Acanthamoeba polyphaga only when used in conjunction with antimycin A, an inhibitor of the conventional cytochrome respiratory pathway. Transcripts for AcAOX are increased during the encystment process, indicating a possible role for alternative respiration during stress.
Original languageEnglish
Pages (from-to)1417-1424
JournalInternational Journal for Parasitology
Volume39
Issue number13
DOIs
Publication statusPublished - Nov 2009

Keywords

  • Acanthamoeba
  • Alternative oxidase
  • Antimicrobial
  • Mitochondria

Cite this

Henriquez-Mui, Fiona ; McBride, James ; Campbell, Sara J. ; Ramos, Tania ; Ingram, Paul R. ; Roberts, Fiona ; Tinney, Sinead ; Roberts, Craig W. / Acanthamoeba alternative oxidase genes : Identification, characterisation and potential as antimicrobial targets. In: International Journal for Parasitology. 2009 ; Vol. 39, No. 13. pp. 1417-1424.
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Acanthamoeba alternative oxidase genes : Identification, characterisation and potential as antimicrobial targets. / Henriquez-Mui, Fiona; McBride, James; Campbell, Sara J.; Ramos, Tania; Ingram, Paul R.; Roberts, Fiona; Tinney, Sinead; Roberts, Craig W.

In: International Journal for Parasitology, Vol. 39, No. 13, 11.2009, p. 1417-1424.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Acanthamoeba alternative oxidase genes

T2 - Identification, characterisation and potential as antimicrobial targets

AU - Henriquez-Mui, Fiona

AU - McBride, James

AU - Campbell, Sara J.

AU - Ramos, Tania

AU - Ingram, Paul R.

AU - Roberts, Fiona

AU - Tinney, Sinead

AU - Roberts, Craig W.

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N2 - Alternative oxidase (AOX) is a mitochondrial protein that acts as an alternative terminal oxidase to the conventional cytochrome oxidases. It is present in certain prokaryotes, plants, fungi and some protozoa but absent in mammals. AOX activity has previously been described in Acanthamoeba, although no genetic evidence has been reported. Herein, two AOX (AcAOX) genes designated isoforms A and B, were obtained from Acanthamoeba castellanii by a combination of degenerate PCR from cDNA and a series of 51 and 3' rapid amplification of cDNA ends. The corresponding genomic sequences of these AcAOXs were also obtained. Each gene spans six exons over a region of 1607 and 1619 bp, respectively. Isoforms A and B have open reading frames of 1113 and 1125 bp, respectively. Each encodes a protein with a predicted molecular weight of 42 kDa. Each AcAOX protein has a predicted cleavable mitochondrial targeting sequence. The full-length AcAOX is functionally active as it complements hemL-deficient Escherichia coli and inhibited by the inhibitor of ACX, salicylhydroxamic acid (SHAM). SHAM is effective against A. castellanii and Acanthamoeba polyphaga only when used in conjunction with antimycin A, an inhibitor of the conventional cytochrome respiratory pathway. Transcripts for AcAOX are increased during the encystment process, indicating a possible role for alternative respiration during stress.

AB - Alternative oxidase (AOX) is a mitochondrial protein that acts as an alternative terminal oxidase to the conventional cytochrome oxidases. It is present in certain prokaryotes, plants, fungi and some protozoa but absent in mammals. AOX activity has previously been described in Acanthamoeba, although no genetic evidence has been reported. Herein, two AOX (AcAOX) genes designated isoforms A and B, were obtained from Acanthamoeba castellanii by a combination of degenerate PCR from cDNA and a series of 51 and 3' rapid amplification of cDNA ends. The corresponding genomic sequences of these AcAOXs were also obtained. Each gene spans six exons over a region of 1607 and 1619 bp, respectively. Isoforms A and B have open reading frames of 1113 and 1125 bp, respectively. Each encodes a protein with a predicted molecular weight of 42 kDa. Each AcAOX protein has a predicted cleavable mitochondrial targeting sequence. The full-length AcAOX is functionally active as it complements hemL-deficient Escherichia coli and inhibited by the inhibitor of ACX, salicylhydroxamic acid (SHAM). SHAM is effective against A. castellanii and Acanthamoeba polyphaga only when used in conjunction with antimycin A, an inhibitor of the conventional cytochrome respiratory pathway. Transcripts for AcAOX are increased during the encystment process, indicating a possible role for alternative respiration during stress.

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KW - Antimicrobial

KW - Mitochondria

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