Amplification of an alternate transporter gene suppresses the avirulent phenotype of glucose transporter null mutants in Leishmania mexicana

Xiuhong Feng, Dayana Rodriguez-Contreras, Cosmo Buffalo, H. G Archie Bouwer, Elizabeth Kruvand, Stephen M. Beverley, Scott Landfear

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

A glucose transporter null mutant of the parasitic protozoan Leishmania mexicana, in which three linked glucose transporter genes have been deleted by targeted gene replacement, is unable to replicate as amastigote forms within phagolysomes of mammalian host macrophages and is avirulent. Spontaneous suppressors of the null mutant have been isolated that partially restore replication of parasites within macrophages. These suppressor mutants have amplified the gene for an alternative hexose transporter, the LmGT4 permease (previously called the D2 permease), on a circular extrachromosomal element, and they overexpress LmGT4 mRNA and protein. The suppressors have also regained the ability to transport hexoses, and they have reverted other phenotypes of the null mutant exhibiting enhanced resistance to oxidative killing, heat shock and starvation for nutrients, as well as augmented levels of the storage carbohydrate β-mannan, increased cell size and increased growth as insect stage promastigotes compared with the unsuppressed mutant. Complementation of the null mutant with the LmGT4 gene on a multicopy episomal expression vector also reverted these phenotypes, confirming that suppression results from amplification of the LmGT4 gene. These results underscore the importance of hexose transporters for the infectious stage of the parasite life cycle.

Original languageEnglish (US)
Pages (from-to)369-381
Number of pages13
JournalMolecular Microbiology
Volume71
Issue number2
DOIs
StatePublished - Jan 2009

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Leishmania mexicana
Facilitative Glucose Transport Proteins
Monosaccharide Transport Proteins
Phenotype
Membrane Transport Proteins
Genes
Parasites
Macrophages
Cell Enlargement
Mannans
Hexoses
Gene Amplification
Starvation
Life Cycle Stages
Insects
Shock
Hot Temperature
Carbohydrates
Food
Messenger RNA

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Amplification of an alternate transporter gene suppresses the avirulent phenotype of glucose transporter null mutants in Leishmania mexicana. / Feng, Xiuhong; Rodriguez-Contreras, Dayana; Buffalo, Cosmo; Bouwer, H. G Archie; Kruvand, Elizabeth; Beverley, Stephen M.; Landfear, Scott.

In: Molecular Microbiology, Vol. 71, No. 2, 01.2009, p. 369-381.

Research output: Contribution to journalArticle

Feng, Xiuhong ; Rodriguez-Contreras, Dayana ; Buffalo, Cosmo ; Bouwer, H. G Archie ; Kruvand, Elizabeth ; Beverley, Stephen M. ; Landfear, Scott. / Amplification of an alternate transporter gene suppresses the avirulent phenotype of glucose transporter null mutants in Leishmania mexicana. In: Molecular Microbiology. 2009 ; Vol. 71, No. 2. pp. 369-381.
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