Helen P. Price

Many malignancies of mature B cells are characterized by chromosomal translocations involving the immunoglobulin heavy chain (IGH) locus on chromosome 14q32.3 and result in deregulated expression of the translocated oncogene. t(2;14)(p13;q32.3) is a rare event in B-cell malignancies. In contrast, gains and amplifications of the same region of chromosome 2p13 have been reported in 20% of extranodal B-cell non-Hodgkin lymphomas (B-NHL), in follicular and mediastinal B-NHL, and in Hodgkin disease (HD). It has been suggested that REL, an NF-kappaB gene family member, mapping within the amplified region, is the pathologic target. However, by molecular cloning of t(2;14)(p13;q32.3) from 3 cases of aggressive B-cell chronic lymphocytic leukemia (CLL)/immunocytoma, this study has shown clustered breakpoints on chromosome 2p13 immediately upstream of a CpG island located about 300 kb telomeric of REL. This CpG island was associated with a Krüppel zinc finger gene (BCL11A), which is normally expressed at high levels only in fetal brain and in germinal center B-cells. There were 3 major RNA isoforms of BCL11A, differing in the number of carboxy-terminal zinc fingers. All 3 RNA isoforms were deregulated as a consequence of t(2;14)(p13;q32.3). BCL11A was highly conserved, being 95% identical to mouse, chicken, and Xenopus homologues. BCL11A was also highly homologous to another gene (BCL11B) on chromosome 14q32.1. BCL11A coamplified with REL in B-NHL cases and HD lymphoma cell lines with gains and amplifications of 2p13, suggesting that BCL11A may be involved in lymphoid malignancies through either chromosomal translocation or amplification.

The essential role of Toll-like receptors (TLR) in innate immune responses to bacterial pathogens is increasingly recognized, but very little is known about the role of TLRs in host defense against infections with eukaryotic pathogens. For the present study, we investigated whether TLRs contribute to the innate and acquired immune response to infection with the intracellular protozoan parasite Leishmania major. Our results show that TLR4 contributes to the control of parasite growth in both phases of the immune response. We also addressed the mechanism that results in killing or growth of the intracellular parasites. Control of parasite replication correlates with the early induction of inducible nitric oxide synthase in TLR4-competent mice, whereas increased parasite survival in host cells from TLR4-deficient mice correlates with a higher activity of arginase, an enzyme known to promote parasite growth. This is the first study showing that TLR4 contributes to the effective control of Leishmania infection in vivo.

Co-translational modification of eukaryotic proteins by N-myristoylation aids subcellular targeting and protein-protein interactions. The enzyme that catalyzes this process,N-myristoyltransferase (NMT), has been characterized in the kinetoplastid protozoan parasites, Leishmania andTrypanosoma brucei. In Leishmania major, the single copy NMT gene is constitutively expressed in all parasite stages as a 48.5-kDa protein that localizes to both membrane and cytoplasmic fractions. Leishmania NMT myristoylates the target acylated Leishmania protein, HASPA, when both are co-expressed in Escherichia coli. Gene targeting experiments have shown that NMT activity is essential for viability inLeishmania. In addition, overexpression of NMT causes gross changes in parasite morphology, including the subcellular accumulation of lipids, leading to cell death. This phenotype is more extreme than that observed in Saccharomyces cerevisiae, in which overexpression of NMT activity has no obvious effects on growth kinetics or cell morphology. RNA interference assays in T. brucei have confirmed that NMT is also an essential protein in both life cycle stages of this second kinetoplastid species, suggesting that this enzyme may be an appropriate target for the development of anti-parasitic agents. Co-translational modification of eukaryotic proteins by N-myristoylation aids subcellular targeting and protein-protein interactions. The enzyme that catalyzes this process,N-myristoyltransferase (NMT), has been characterized in the kinetoplastid protozoan parasites, Leishmania andTrypanosoma brucei. In Leishmania major, the single copy NMT gene is constitutively expressed in all parasite stages as a 48.5-kDa protein that localizes to both membrane and cytoplasmic fractions. Leishmania NMT myristoylates the target acylated Leishmania protein, HASPA, when both are co-expressed in Escherichia coli. Gene targeting experiments have shown that NMT activity is essential for viability inLeishmania. In addition, overexpression of NMT causes gross changes in parasite morphology, including the subcellular accumulation of lipids, leading to cell death. This phenotype is more extreme than that observed in Saccharomyces cerevisiae, in which overexpression of NMT activity has no obvious effects on growth kinetics or cell morphology. RNA interference assays in T. brucei have confirmed that NMT is also an essential protein in both life cycle stages of this second kinetoplastid species, suggesting that this enzyme may be an appropriate target for the development of anti-parasitic agents. N-myristoyltransferase hydrophilic acylated surface protein glycophosphatidylinositol open reading frame RNA interference ADP-ribosylation factor group of overlapping clones Myristoyl-CoA:protein N-myristoyltransferase (NMT1; EC 2.3.1.97) catalyzes the covalent co-translational attachment of the fatty acid, myristate (C14:0) to the amino-terminal glycine residue of a number of eukaryotic cellular and viral proteins of diverse function (1Gordon J.I. Duronio R.J. Rudnick D.A. Adams S.P. Gokel G.W. J. Biol. Chem. 1991; 266: 8647-8650Abstract Full Text PDF PubMed Google Scholar). These include cAMP-dependent serine/threonine kinases, members of the p60 Src family of tyrosine kinases, retroviral gag polyprotein precursors such as HIV-1pr55, viral capsid components, and the α-subunit of many signal-transducing, heteromeric G proteins. Although some myristoylated proteins are cytosolic, many are associated with cellular membranes where myristoylation facilitates membrane attachment. The addition of myristate can also stabilize protein-protein interactions, and many acylated proteins require this modification for full expression of their biological function (2McIlhinney R.A. Methods Mol. Biol. 1998; 88: 211-225PubMed Google Scholar). Genetic studies have shown that theNMT gene is essential for the survival ofSaccharomyces cerevisiae (3Duronio R.J. Towler D.A. Heuckeroth R.O. Gordon J.I. Science. 1989; 243: 796-800Crossref PubMed Scopus (158) Google Scholar) and the pathogenic fungi,Candida albicans and Cryptococcus neoformans (4Lodge J.K. Johnson R.L. Weinberg R.A. Gordon J.I. J. Biol. Chem. 1994; 269: 2996-3009Abstract Full Text PDF PubMed Google Scholar). Given the critical role of N-myristoylation in the cell, NMT has been developed as a target for the development of antifungal chemotherapeutic agents. The NMT enzyme has an ordered Bi-Bi reaction mechanism (5Rudnick D.A. McWherter C.A. Rocque W.J. Lennon P.J. Getman D.P. Gordon J.I. J. Biol. Chem. 1991; 266: 9732-9739Abstract Full Text PDF PubMed Google Scholar), binding first to myristoyl-CoA, with the resulting conformational changes generating a peptide-binding site. Subsequent formation of a ternary myristoyl-CoA:NMT-peptide complex leads to catalysis and product release. The myristoyl-CoA binding sites of purified human NMT and the fungal NMTs are highly conserved, but their peptide binding specificities are divergent (6Johnson D.R. Bhatnagar R.S. Knoll L.J. Gordon J.I. Annu. Rev. Biochem. 1994; 63: 869-914Crossref PubMed Scopus (371) Google Scholar). By exploiting these differences, peptide and peptidomimetic inhibitors with selectivity for fungal NMTs have been designed. For example, an inhibitor based on the N terminus of ADP-ribosylation factor (ARF), the major myristoylated protein detected in [3H]myristate-labeled C. neoformans (causative agent of chronic meningitis in AIDS patients), is competitive for peptide and noncompetitive for myristoyl-CoA binding (7Langner C.A. Lodge J.K. Travis S.J. Caldwell J.E. Lu T. Li Q. Bryant M.L. Devadas B. Gokel G.W. Kobayashi G.S. J. Biol. Chem. 1992; 267: 17159-17169Abstract Full Text PDF PubMed Google Scholar), and a nonpeptide inhibitor based on this sequence is at least ∼5-fold selective for C. neoformansagainst human NMT (8Lodge J.K. Jackson-Machelski E. Higgins M. McWherter C.A. Sikorski J.A. Devadas B. Gordon J.I. J. Biol. Chem. 1998; 273: 12482-12491Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar). Similarly, peptidomimetic inhibitors based on the N-terminal sequence of C. albicans ARF are 560-fold selective for the fungal NMT as compared with the human enzyme. Gel mobility assays indicate that a single dose of 200 μm is sufficient to produce ∼50% reduction in the myristoylation ofC. albicans ARF, which is consistent with fungistatic activity (9Lodge J.K. Jackson-Machelski E. Devadas B. Zupec M.E. Getman D.P. Kishore N. Freeman S.K. McWherter C.A. Sikorski J.A. Gordon J.I. Microbiology. 1997; 143: 357-366Crossref PubMed Scopus (35) Google Scholar). Recent elucidation of the S. cerevisiae and C. albicans NMT crystal structures (10Weston S.A. Camble R. Colls J. Rosenbrock G. Taylor I. Egerton M. Tucker A.D. Tunnicliffe A. Mistry A. Mancia F. de la Fortelle E. Irwin J. Bricogne G. Pauptit R.A. Nat. Struct. Biol. 1998; 5: 213-221Crossref PubMed Scopus (103) Google Scholar, 11Bhatnagar R.S. Futterer K. Waksman G. Gordon J.I. Biochim. Biophys. Acta. 1999; 1441: 162-172Crossref PubMed Scopus (57) Google Scholar, 12Farazi T.A. Waksman G. Gordon J.I. J. Biol. Chem. 2001; 276: 39501-39504Abstract Full Text Full Text PDF PubMed Scopus (434) Google Scholar) will facilitate a better understanding of the inhibition mechanisms of these enzymes. Parasitic kinetoplastid protozoa of the species Leishmaniaand Trypanosoma are causative agents of some of the most debilitating tropical infections currently affecting world populations (see, on the World Wide Web, www.who.int/tdr/index.html). Metabolic labeling studies have identified at least ten [3H]myristate-labeled proteins in Leishmania major, including the ARF-like protein 1 (53Menon M.R. Molecular Cloning and Characterisation of Leishmania major N-myristoyltransferase-a Putative Drug TargetPh.D. thesis. University of London, 2002Google Scholar). and the infective stage specific, hydrophilic acylated surface proteins (HASPs) (13Flinn H.M. Rangarajan D. Smith D.F. Mol. Biochem. Parasitol. 1994; 65: 259-270Crossref PubMed Scopus (64) Google Scholar, 14McKean P.G. Delahay R. Pimenta P.F. Smith D.F. Mol. Biochem. Parasitol. 1997; 85: 221-231Crossref PubMed Scopus (31) Google Scholar). HASPB, a candidate vaccine for visceral leishmaniasis (15Stager S. Smith D.F. Kaye P.M. J. Immunol. 2000; 165: 7064-7071Crossref PubMed Scopus (163) Google Scholar), requires N-terminal myristoylation and palmitoylation for translocation to the parasite plasma membrane (16Denny P.W. Gokool S. Russell D.G. Field M.C. Smith D.F. J. Biol. Chem. 2000; 275: 11017-11025Abstract Full Text Full Text PDF PubMed Scopus (143) Google Scholar). Myristate is also a component of the glycophosphatidylinositol (GPI) lipid anchors that tether the major classes of surface molecules in trypanosomatid parasites. InLeishmania, the myristate-containing GPI anchors of the surface glycoinositolphospholipids are remodeled during synthesis by a reaction involving myristate exchange from a myristoyl-CoA donor (17Ralton J.E. McConville M.J. J. Biol. Chem. 1998; 273: 4245-4257Abstract Full Text Full Text PDF PubMed Scopus (99) Google Scholar). In stages of Trypanosoma fatty is also in synthesis of the surface GPI which in A. J. Biol. Chem. 2000; 275: Full Text Full Text PDF PubMed Scopus Google Scholar). a myristate are in T. to a on GPI and protein synthesis J. Adams S.P. Gordon J.I. G.W. Science. 1991; PubMed Scopus Google Scholar, Lu T. Gokel G.W. G.W. Gordon J.I. S. A. 1994; PubMed Scopus Google Scholar). that myristate inhibitors of are also to Leishmania and the of NMT in both major brucei. with the pathogenic expression of NMT is essential for viability in these parasites, of enzyme activity parasite to modification of target proteins. in as (13Flinn H.M. Rangarajan D. Smith D.F. Mol. Biochem. Parasitol. 1994; 65: 259-270Crossref PubMed Scopus (64) Google Scholar). at with inhibitors in as that been to as (16Denny P.W. Gokool S. Russell D.G. Field M.C. Smith D.F. J. Biol. Chem. 2000; 275: 11017-11025Abstract Full Text Full Text PDF PubMed Scopus (143) Google Scholar). to to the the in for all including with morphology. in and the of the NMT gene from major at and the and based on the NMT and on the World Wide at and to major The and to a major A. H.M. Smith D.F. 1998; PubMed Scopus Google Scholar). the clones to the sequence on major NMT from A. H.M. Smith D.F. 1998; PubMed Scopus Google Scholar) as and The to and sequence of the NMT from Leishmania number Leishmania number The T. brucei NMT gene identified as of a on in at of the Trypanosoma NMT is also number of the T. brucei open reading frame The major from and The major from P.G. Delahay R. Pimenta P.F. Smith D.F. Mol. Biochem. Parasitol. 1997; 85: 221-231Crossref PubMed Scopus (31) Google Scholar) and The and the of both as For expression in Escherichia the NMT with and the expression to The with and the expression generating The expressed from this has an N-terminal myristoylation of major NMT by in E. and the in to protein purified by and for and of For in proteins by and the resulting with purified NMT P.G. Delahay R. Pimenta P.F. Smith D.F. Mol. Biochem. Parasitol. 1997; 85: 221-231Crossref PubMed Scopus (31) Google Scholar), M.J. J. PubMed Google Scholar), of E. P.G. Smith D.F. Biochem. J. 2001; PubMed Scopus Google Scholar), and C. K. K. Mol. Biochem. Parasitol. 2001; PubMed Scopus Google Scholar). detected an This based on the in R.A. Methods Mol. Biol. 1998; 88: 211-225PubMed Google E. with and or in to in and and expression of proteins by the addition of 1 the with of in of to and of to the The at for a to and by proteins detected by at and membrane and cytoplasmic as E. P.G. Smith D.F. Biochem. J. 2001; PubMed Scopus Google Scholar). in both by and as target both NMT a gene or a gene with and of the NMT These based on the A. S. A. 1991; 88: PubMed Scopus Google Scholar), and a of the NMT and Cloning sites are shown in and major sequence is shown in The product with and generating a of the NMT the and and the sites of generating the gene from J.E. R. M.J. S. A. PubMed Scopus Google Scholar) as P.G. P.W. E. J.K. Smith D.F. 2001; PubMed Scopus Google Scholar) and to the gene in targeting by with and with purified and to as Mol. Biol. PubMed Scopus Google Scholar). in the of and clones and for as P.G. P.W. E. J.K. Smith D.F. 2001; PubMed Scopus Google Scholar). The and by and with N-terminal by with proteins detected as For of major or overexpression in parasites, to the the expression H.M. G. 1992; PubMed Scopus Google Scholar), generating NMT with of to in the of and as and in P.G. P.W. E. J.K. Smith D.F. 2001; PubMed Scopus Google the NMT from T. brucei the and T. brucei sequence is shown in and sites are The with and the sites of the RNA interference which expression of RNA from S. J.E. Mol. Biochem. Parasitol. 2000; PubMed Scopus Google Scholar), generating The T. brucei E. S. C. Mol. Biochem. Parasitol. 1999; PubMed Scopus Google Scholar), the RNA and in at in with and with of as Russell D.G. J.E. J. 1999; PubMed Google Scholar). The to and and for cell been The expression of RNA by the addition of 1 to to 1 in a at and by as The T. brucei E. S. C. Mol. Biochem. Parasitol. 1999; PubMed Scopus Google Scholar) at with in with and with as S. C. Mol. Biochem. Parasitol. 1997; 85: PubMed Scopus Google Scholar). The to for the addition of The to and for cell been of RNA by the addition of 1 to to in as in and at 1 in and in in in with acid, an in at the and in on a with and and on a For lipid at to the to on for as a at the stage during and with and as myristate are competitive inhibitors of first these for their on Leishmania growth and in the myristate and with a the to growth in the and parasite a by on the inhibitor reduction in parasite compared with with both inhibitors a the with and when compared with the These as with T. myristate are to Leishmania most to effects on GPI and as as N-myristoylation of target proteins. to the of viability observed in the of these with N-myristoylation of the single characterized Leishmania protein, HASPB, is expressed in the growth of to infective (13Flinn H.M. Rangarajan D. Smith D.F. Mol. Biochem. Parasitol. 1994; 65: 259-270Crossref PubMed Scopus (64) Google Scholar, 14McKean P.G. Delahay R. Pimenta P.F. Smith D.F. Mol. Biochem. Parasitol. 1997; 85: 221-231Crossref PubMed Scopus (31) Google Scholar). The major NMT gene by of by sequence from the gene a for a 48.5-kDa protein of number This protein with NMT from species, as shown in the sequence in major NMT and with S. C. The NMTs of species are highly with major enzyme and in the the in the trypanosomatid species, T. the to of NMT of brucei with major with all NMTs the major and T. brucei are divergent at their N are in and N-terminal have been in targeting of human and R.L. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar). many of the fungal NMT to be in the enzyme mechanism or to have critical (6Johnson D.R. Bhatnagar R.S. Knoll L.J. Gordon J.I. Annu. Rev. Biochem. 1994; 63: 869-914Crossref PubMed Scopus (371) Google Scholar) are in major brucei. is of S. cerevisiae NMT in fungal of this residue has a selective on the S. peptide binding Jackson-Machelski E. Gordon J.I. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In major and T. brucei this residue is by of the C. albicans enzyme has that and the of the of the enzyme (10Weston S.A. Camble R. Colls J. Rosenbrock G. Taylor I. Egerton M. Tucker A.D. Tunnicliffe A. Mistry A. Mancia F. de la Fortelle E. Irwin J. Bricogne G. Pauptit R.A. Nat. Struct. Biol. 1998; 5: 213-221Crossref PubMed Scopus (103) Google Scholar). These are in both T. brucei but is also an of in and to the first of these in the parasite of is in T. brucei These that the parasite NMTs and structures that their to the human and fungal enzymes. major NMT is a single copy gene by and Similarly, the of the World species, and T. brucei also Leishmania cycle the and of the NMT expression during the major life of or and infective and from and by NMT as a single 48.5-kDa protein, in all life cycle stages In the expression of HASPB, a of the hydrophilic acylated surface protein to infective parasite stages as (13Flinn H.M. Rangarajan D. Smith D.F. Mol. Biochem. Parasitol. 1994; 65: 259-270Crossref PubMed Scopus (64) Google Scholar, 14McKean P.G. Delahay R. Pimenta P.F. Smith D.F. Mol. Biochem. Parasitol. 1997; 85: 221-231Crossref PubMed Scopus (31) Google Scholar). of the protein, as a for NMT is constitutively expressed NMT has been to the in S. cerevisiae and to be associated with cellular membranes L.J. Gordon J.I. J. Biol. Chem. 1992; 267: Full Text PDF PubMed Google Scholar). a of NMT has been shown to be M. Egerton M. J. 1997; PubMed Google Scholar), and NMTs in R.A. K. PubMed Scopus Google Scholar, Mol. Biochem. PubMed Scopus Google Scholar) are all associated with as membrane proteins. This is also the for major NMT of parasite membrane and cytoplasmic with that of major enzyme is associated with for these major surface protein detected in the membrane as a hydrophilic protein that with membranes in E. P.G. Smith D.F. Biochem. J. 2001; PubMed Scopus Google Scholar), detected in the cytoplasmic (2McIlhinney R.A. Methods Mol. Biol. 1998; 88: 211-225PubMed Google Scholar) to the activity of major NMT in In these a major NMT and a the of an NMT coli. expression with by number have been in these including and HASPA, a second of the hydrophilic acylated surface protein In the in E. with and to labeling with The by by and of protein expression in the proteins at and expressed in cell These identified as NMT and HASPA, by proteins when in the The is to a complex with myristoyl-CoA (5Rudnick D.A. McWherter C.A. Rocque W.J. Lennon P.J. Getman D.P. Gordon J.I. J. Biol. Chem. 1991; 266: 9732-9739Abstract Full Text PDF PubMed Google Scholar), the of the 48.5-kDa The at myristoylated by the NMT expressed from This labeling is to with 1 of an with no of protein expression and of NMT in the of in myristoylation of an E. protein have been observed R.J. Rudnick D.A. Johnson R.L. Johnson D.R. Gordon J.I. J. Biol. 1991; PubMed Scopus Google Scholar, R.J. Rudnick D.A. Adams S.P. Towler D.A. Gordon J.I. J. Biol. Chem. 1991; 266: Full Text PDF PubMed Google Scholar). these major NMT is and is for the N-myristoylation of the of NMT expression in life gene targeting to or both of the or by from the for expression K. PubMed Scopus Google Scholar), to the NMT and for In these of clones for clones and from NMT a second of to clones that to both of these the of clones by and of these clones by to NMT when of the NMT gene parasites, to targeting the second for clones in this both NMT of a of clones is shown in B. of the the NMT is detected on and in clones and In clones and of the NMT is detected by of a to of and The second NMT gene has been by gene targeting in in which NMT are as the are The expression of NMT in the clones as a protein, the NMT expression as compared with and for clones of NMT in protein the of to the gene copy number on the of the clones also a clones a in growth compared with parasite in these by when to both of essential proteins N. E. Smith D.F. Parasitol. 1999; PubMed Scopus Google Scholar, R. S. A. PubMed Scopus Google Scholar, Mol. PubMed Scopus Google Scholar). a that NMT is an essential gene the effects of NMT expression on parasite the NMT The resulting parasites, by their to and developed during growth on and in These with subcellular and of membranes with in and of as a which to be by with confirmed that these structures are in of parasite from NMT at the as for a in the expression of NMT protein compared with the clones NMT viability for a but cell in all compared with clones with the This is a more extreme phenotype than that in in which with a in NMT expression but a growth a in NMT expression is an to is the on in by in NMT expression and changes to shown in for expression of are development has been as a for gene and in T. brucei S. J.E. Mol. Biochem. Parasitol. 2000; PubMed Scopus Google Scholar, A. T. E. C. E. 2000; PubMed Scopus Google Scholar, E. A. C. R. Biol. PubMed Scopus Google Scholar). to the phenotype of in to target expression from the T. brucei NMT gene in both and parasite with of of the of the RNA and and the a and at for the first when the that growth of the the as this the parasites, a more growth to the 1 cell at parasite at a in NMT protein expression by with the at which the NMT expression be detected by by which a reduction in parasite growth and in cell This at with parasite viability and no NMT an accumulation of with morphology, including or more than and plasma membrane and This of be to the effects of NMT on modification of a of target proteins. of of the protein C. K. K. Mol. Biochem. Parasitol. 2001; PubMed Scopus Google Scholar), during the of this when growth of of the and clones compared The for and to leading to a of cell by can as major, NMT protein is essential for viability of T. brucei in both life cycle stages of the The of N-myristoylation in the synthesis of with a role in or cellular has on the enzyme that catalyzes this The development of peptide and peptidomimetic for fungal has been by of the of the enzyme. that both and inhibition of NMT activity can be when compared with enzyme activity (8Lodge J.K. Jackson-Machelski E. Higgins M. McWherter C.A. Sikorski J.A. Devadas B. Gordon J.I. J. Biol. Chem. 1998; 273: 12482-12491Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar, 12Farazi T.A. Waksman G. Gordon J.I. J. Biol. Chem. 2001; 276: 39501-39504Abstract Full Text Full Text PDF PubMed Scopus (434) Google Scholar). this have and characterized NMT from trypanosomatid parasite species, in to the in these and to the of NMT as a target in the that The major and T. brucei NMTs with characterized NMTs in their including of essential for including the identified at the (10Weston S.A. Camble R. Colls J. Rosenbrock G. Taylor I. Egerton M. Tucker A.D. Tunnicliffe A. Mistry A. Mancia F. de la Fortelle E. Irwin J. Bricogne G. Pauptit R.A. Nat. Struct. Biol. 1998; 5: 213-221Crossref PubMed Scopus (103) Google Scholar) and the and in S. critical for myristoyl-CoA binding and product R.S. Futterer K. Waksman G. Gordon J.I. Biochim. Biophys. Acta. 1999; 1441: 162-172Crossref PubMed Scopus (57) Google Scholar). The N of the parasite sequence and the in that are also from the NMTs of eukaryotic species to In this the of than co-translational N-myristoylation of the protein, that NMT is to more than J. S. M.C. S.J. Science. 2000; PubMed Scopus Google Scholar), at least in This with the of a second NMT from a second and the expression of NMT in J. Biol. Chem. 1998; 273: Full Text Full Text PDF PubMed Scopus Google Scholar). Leishmania by a single copy is membranes and the NMT is constitutively expressed in Leishmania species as in the pathogenic major NMT in E. has a for enzyme activity in as with S. cerevisiae which has fatty and peptide specificities from of the enzyme when expressed in R.J. Rudnick D.A. Adams S.P. Towler D.A. Gordon J.I. J. Biol. Chem. 1991; 266: Full Text PDF PubMed Google Scholar). with S. cerevisiae major species are detected Leishmania NMT and a the complex and the myristoylated of these are when the NMT is most to of the with this of a of major NMT based on the S. has shown that this is on the surface of the protein and the K. R. C. Smith D.F. Mol. Biochem. Parasitol. Scholar). with purified enzyme will the of this and and The and of myristate inhibitors of have been (7Langner C.A. Lodge J.K. Travis S.J. Caldwell J.E. Lu T. Li Q. Bryant M.L. Devadas B. Gokel G.W. Kobayashi G.S. J. Biol. Chem. 1992; 267: 17159-17169Abstract Full Text PDF PubMed Google Scholar, Lu T. Gokel G.W. G.W. Gordon J.I. S. A. 1994; PubMed Scopus Google Scholar, S.A. R.L. PubMed Scopus Google Scholar), these The the growth of major to effects on cellular peptide binding inhibitors will be to this and growth inhibition with NMT an have confirmed that NMT expression is essential for the viability of both Leishmania and T. parasites, NMT protein also have In the accumulation of lipid in Leishmania NMT to cell may be a phenotype to that of surface By overexpression of NMT activity in S. cerevisiae has no on growth kinetics or cell (3Duronio R.J. Towler D.A. Heuckeroth R.O. Gordon J.I. Science. 1989; 243: 796-800Crossref PubMed Scopus (158) Google Scholar). N-myristoylation of to this parasite at the of for the which include GPI (17Ralton J.E. McConville M.J. J. Biol. Chem. 1998; 273: 4245-4257Abstract Full Text Full Text PDF PubMed Scopus (99) Google Scholar). the of de myristate synthesis in T. brucei D. Parasitol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar) that fatty may also and that myristate may be a factor to the accumulation of lipid experiments are to the of NMT expression in of Leishmania and to the that to the of viability is that of the proteins in cell function are as as P.G. P.W. E. J.K. Smith D.F. 2001; PubMed Scopus Google Scholar). By ARF myristoylation has been shown to growth in C. albicans and S. cerevisiae (8Lodge J.K. Jackson-Machelski E. Higgins M. McWherter C.A. Sikorski J.A. Devadas B. Gordon J.I. J. Biol. Chem. 1998; 273: 12482-12491Abstract Full Text Full Text PDF PubMed Scopus (42) Google Scholar, J.K. Jackson-Machelski E. Devadas B. Zupec M.E. Getman D.P. Kishore N. Freeman S.K. McWherter C.A. Sikorski J.A. Gordon J.I. Microbiology. 1997; 143: 357-366Crossref PubMed Scopus (35) Google Scholar), suggesting that a of this protein family may be for viability in trypanosomatid parasites. the myristoylated proteins the of trypanosomatid including their single gene copy and expression in parasite that these may be appropriate for the development of from the and for parasite and for with the T. brucei and T. for to sequence and for and for with T. brucei and and members of the Smith for