Remodeling of protein and mRNA expression in Leishmania mexicana induced by deletion of glucose transporter genes

Xiuhong Feng, Torben Feistel, Cosmo Buffalo, Ashley McCormack, Elizabeth Kruvand, Dayana Rodriguez-Contreras, Natalia S. Akopyants, P. K. Umasankar, Larry David, Armando Jardim, Stephen M. Beverley, Scott Landfear

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Glucose is a major nutrient in the insect vector stage of Leishmania parasites. Glucose transporter null mutants of Leishmania mexicana exhibit profound phenotypic changes in both insect stage promastigotes and mammalian host stage amastigotes that reside within phagolysosomes of host macrophages. Some of these phenotypic changes could be either mediated or attenuated by changes in gene expression that accompany deletion of the glucose transporter genes. To search for changes in protein expression, the profile of proteins detected on two-dimensional gels was compared for wild type and glucose transporter null mutant promastigotes. A total of 50 spots whose intensities changed significantly and consistently in multiple experiments were detected, suggesting that a cohort of proteins is altered in expression levels in the null mutant parasites. Following identification of proteins by mass spectrometry, 3 such regulated proteins were chosen for more detailed analysis: mitochondrial aldehyde dehydrogenase, ribokinase, and hexokinase. Immunoblots employing antisera against these enzymes confirmed that their levels were upregulated, both in glucose transporter null mutants and in wild type parasites starved for glucose. Quantitative reverse transcriptase PCR (qRT-PCR) revealed that the levels of mRNAs encoding these enzymes were also enhanced. Global expression profiling using microarrays revealed a limited number of additional changes, although the sensitivity of the microarrays to detect modest changes in amplitude was less than that of two-dimensional gels. Hence, there is likely to be a network of proteins whose expression levels are altered by genetic ablation of glucose transporters, and much of this regulation may be reflected by changes in the levels of the cognate mRNAs. Some of these changes in protein expression may reflect an adaptive response of the parasites to limitation of glucose.

Original languageEnglish (US)
Pages (from-to)39-48
Number of pages10
JournalMolecular and Biochemical Parasitology
Volume175
Issue number1
DOIs
StatePublished - Jan 2011

Fingerprint

Leishmania mexicana
Facilitative Glucose Transport Proteins
Messenger RNA
Parasites
Genes
Proteins
Glucose
Gels
Insect Vectors
Phagosomes
Hexokinase
Leishmania
Enzymes
Reverse Transcriptase Polymerase Chain Reaction
Insects
Immune Sera
Mass Spectrometry
Macrophages
Gene Expression
Food

Keywords

  • Glucose starvation
  • Glucose transporter null mutant
  • Leishmania mexicana
  • Regulation of protein expression

ASJC Scopus subject areas

  • Molecular Biology
  • Parasitology

Cite this

Remodeling of protein and mRNA expression in Leishmania mexicana induced by deletion of glucose transporter genes. / Feng, Xiuhong; Feistel, Torben; Buffalo, Cosmo; McCormack, Ashley; Kruvand, Elizabeth; Rodriguez-Contreras, Dayana; Akopyants, Natalia S.; Umasankar, P. K.; David, Larry; Jardim, Armando; Beverley, Stephen M.; Landfear, Scott.

In: Molecular and Biochemical Parasitology, Vol. 175, No. 1, 01.2011, p. 39-48.

Research output: Contribution to journalArticle

Feng, X, Feistel, T, Buffalo, C, McCormack, A, Kruvand, E, Rodriguez-Contreras, D, Akopyants, NS, Umasankar, PK, David, L, Jardim, A, Beverley, SM & Landfear, S 2011, 'Remodeling of protein and mRNA expression in Leishmania mexicana induced by deletion of glucose transporter genes', Molecular and Biochemical Parasitology, vol. 175, no. 1, pp. 39-48. https://doi.org/10.1016/j.molbiopara.2010.08.008
Feng, Xiuhong ; Feistel, Torben ; Buffalo, Cosmo ; McCormack, Ashley ; Kruvand, Elizabeth ; Rodriguez-Contreras, Dayana ; Akopyants, Natalia S. ; Umasankar, P. K. ; David, Larry ; Jardim, Armando ; Beverley, Stephen M. ; Landfear, Scott. / Remodeling of protein and mRNA expression in Leishmania mexicana induced by deletion of glucose transporter genes. In: Molecular and Biochemical Parasitology. 2011 ; Vol. 175, No. 1. pp. 39-48.
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