Myristoyl-CoA:protein N-myristoyltransferase depletion in trypanosomes causes avirulence and endocytic defects

TY - JOUR

T1 - Myristoyl-CoA:protein N-myristoyltransferase depletion in trypanosomes causes avirulence and endocytic defects

AU - Price, Helen P.

AU - Guether, M. Lucia S.

AU - Ferguson, Michael A. J.

AU - Smith, Deborah F.

PY - 2010/1

Y1 - 2010/1

N2 - The enzyme myristoyl-CoA: protein N-myristoyltransferase (NMT)catalyses the co-translational covalent attachment of the fatty acid myristate to the N-terminus of target proteins. NMT is known to be essential for viability in Trypanosoma brucei and Leishmania major. Here we describe phenotypic analysis of T. brucei bloodstream form cells following knockdown of NMT expression by tetracycline-inducible RNA interference. Cell death occurs from 72 h post-induction, with approximately 50% of cells displaying a defect in endocytic uptake by this time. The majority of these induced cells do not have an enlarged flagellar pocket typical of a block in endocytosis but vesicle accumulation around the flagellar pocket indicates a defect in vesicular progression following endocytic fusion. Induced parasites have a wild-type or slightly enlarged Golgi apparatus, unlike the phenotype of cells with reduced expression of a major N-myristoylated protein, ARL1. Critically we show that following NMT knockdown, T. brucei bloodstream form cells are unable to establish an infection in a mouse model, therefore providing further validation of this enzyme as a target for drug development. (C) 2009 Elsevier B.V. All rights reserved.

AB - The enzyme myristoyl-CoA: protein N-myristoyltransferase (NMT)catalyses the co-translational covalent attachment of the fatty acid myristate to the N-terminus of target proteins. NMT is known to be essential for viability in Trypanosoma brucei and Leishmania major. Here we describe phenotypic analysis of T. brucei bloodstream form cells following knockdown of NMT expression by tetracycline-inducible RNA interference. Cell death occurs from 72 h post-induction, with approximately 50% of cells displaying a defect in endocytic uptake by this time. The majority of these induced cells do not have an enlarged flagellar pocket typical of a block in endocytosis but vesicle accumulation around the flagellar pocket indicates a defect in vesicular progression following endocytic fusion. Induced parasites have a wild-type or slightly enlarged Golgi apparatus, unlike the phenotype of cells with reduced expression of a major N-myristoylated protein, ARL1. Critically we show that following NMT knockdown, T. brucei bloodstream form cells are unable to establish an infection in a mouse model, therefore providing further validation of this enzyme as a target for drug development. (C) 2009 Elsevier B.V. All rights reserved.

KW - Myristoyl-CoA:protein

KW - N-myristoyltransferase

KW - N-Myristoylation

KW - Trypanosoma brucei

KW - Endocytosis

KW - RNA interference

KW - Drug target

KW - CANDIDA-ALBICANS

KW - ESSENTIAL ENZYME

KW - BRUCEI

KW - PROTEINS

KW - COA

KW - N-MYRISTOYLTRANSFERASE-1

KW - LEISHMANIA

KW - VIABILITY

UR - http://www.scopus.com/inward/record.url?scp=71549117083&partnerID=8YFLogxK

U2 - 10.1016/j.molbiopara.2009.09.006

DO - 10.1016/j.molbiopara.2009.09.006

M3 - Article

C2 - 19782106

SN - 0166-6851

VL - 169

SP - 55

EP - 58

JO - MOLECULAR AND BIOCHEMICAL PARASITOLOGY

JF - MOLECULAR AND BIOCHEMICAL PARASITOLOGY

IS - 1

ER -