Down-regulation of MARCKS-related Protein (MRP) in Macrophages Infected with Leishmania

Corradin, Sally Betz; Mauël, Jacques; Ransijn, Adriana; Stürzinger, Christoph; Vergères, Guy

doi:10.1074/jbc.274.24.16782

Cited by 37 publications

(33 citation statements)

References 46 publications

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“…SDS-PAGE and Western Blot Analysis-Western blot analysis was performed as described previously (14). The anti-MRP antibody used recognizes both myristoylated and unmyristoylated MRPs as well as MRP phosphorylated in vitro by the catalytic subunit of PKC (25).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

“…We recently suggested that MRP levels might also be regulated by proteolysis under certain conditions (14). Activation of murine macrophages by bacterial lipopolysaccharide or cytokines strongly up-regulates MRP mRNA and protein expression (5,14).…”

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confidence: 99%

“…Activation of murine macrophages by bacterial lipopolysaccharide or cytokines strongly up-regulates MRP mRNA and protein expression (5,14). However, infection of macrophages with promastigotes of the obligate intracellular parasite Leishmania at the time of activation or up to 48 h post-activation results in significant MRP depletion (14). Moreover, experiments with [ 3 H]myristate-labeled macrophages (14) and Western blot analysis of macrophage lysates using an antibody directed against the C terminus of MRP 2 revealed the appearance of lower molecular weight MRP fragments in Leishmania-infected cells.…”

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confidence: 99%

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MARCKS-related Protein (MRP) Is a Substrate for the Leishmania major Surface Protease Leishmanolysin (gp63)

Corradin¹,

Ransijn²,

Corradin³

et al. 1999

Journal of Biological Chemistry

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Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP; MacMARCKS) are protein kinase C substrates in diverse cell types. Activation of murine macrophages by cytokines increases MRP expression, but infection with Leishmania promastigotes during activation results in MRP depletion. We therefore examined the effect of Leishmania major LV39 on recombinant MRP. Both live promastigotes and a soluble fraction of LV39 lysates degraded MRP to yield lower molecular weight fragments. Degradation was independent of MRP myristoylation and was inhibited by protein kinase C-dependent phosphorylation of MRP. MRP was similarly degraded by purified leishmanolysin (gp63), a Leishmania surface metalloprotease. Degradation was evident at low enzyme/substrate ratios, over a broad pH range, and was inhibited by 1,10-phenanthroline and by a hydroxamate dipeptide inhibitor of leishmanolysin. Using mass spectrometric analysis, cleavage was shown to occur within the effector domain of MRP between Ser 92 and Phe 93, in accordance with the substrate specificity of leishmanolysin. Moreover, an MRP construct in which the effector domain had been deleted was resistant to cleavage. Thus, Leishmania infection may result in leishmanolysin-dependent hydrolysis of MRP, a major protein kinase C substrate in macrophages.Myristoylated alanine-rich C kinase substrate (MARCKS) 1 and MARCKS-related protein (MRP), also known as Mac-MARCKS, are members of a highly acidic myristoylated family of protein kinase C (PKC) substrates (1, 2). The primary structures of MARCKS and MRP exhibit significant homology, including a highly basic stretch of amino acid residues known as the effector domain (also as the phosphorylation site domain), which contains the serine residues subject to PKC-dependent phosphorylation as well as binding sites for calmodulin and actin. Whereas MARCKS is widely distributed in diverse cell types, MRP is present primarily in brain and reproductive tissue (3, 4) as well as in macrophages, where it was first characterized (5). The biologic functions of MARCKS proteins are unknown. Due to their high effector domain homology, it is also possible that MRP and MARCKS play overlapping roles in some cells. In macrophages, both proteins colocalize in the cytosol in association with components of the actin cytoskeleton (6 -9) and consequently are thought to participate in major cellular responses such as phagocytosis, motility, and membrane trafficking.The expression of MARCKS proteins appears to be highly regulated, and in vitro studies have demonstrated up-or downregulation of MARCKS at both the transcriptional and posttranscriptional levels (5, 10, 11). One mechanism of post-transcriptional regulation involves proteolytic degradation. Spizz and Blackshear (12) recently identified cathepsin B as a cellular MARCKS-cleaving enzyme in fibroblasts. They suggested that cleavage might occur within lysosomes as a result of specific lysosomal targeting sequences identified within the MARCKS primary sequence. At least one c...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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MARCKS-related Protein (MRP) Is a Substrate for the Leishmania major Surface Protease Leishmanolysin (gp63)

Corradin¹,

Ransijn²,

Corradin³

et al. 1999

Journal of Biological Chemistry

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“…73 Briefly, 10 5 BMM were distributed in 24-well cell culture plates, each well containing a round sterile glass coverslip, before infection (5 parasites per BMM). Where indicated, BMM were stimulated with 50 U ml 71 g-IFN and 10 ng ml 71 LPS.…”

Section: Bmm In Vitro Infection and Tunel Assaymentioning

confidence: 99%

Cell death in Leishmania induced by stress and differentiation: programmed cell death or necrosis?

2002

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Unicellular organisms, such as the protozoan parasite Leishmania, can be stimulated to show some morphological and biochemical features characteristic of mammalian apoptosis. This study demonstrates that under a variety of stress conditions such as serum deprivation, heat shock and nitric oxide, cell death can be induced leading to genomic DNA fragmentation into oligonucleosomes. DNA fragmentation was observed, without induction, in the infectious stages of the parasite, and correlated with the presence of internucleosomal nuclease activity, visualisation of 45 to 59 kDa nucleases and detection of TUNEL-positive nuclei. DNA fragmentation was not dependent on active effector downstream caspases nor on the lysosomal cathepsin L-like enyzmes CPA and CPB. These data are consistent with the presence of a caspase-independent cell death mechanism in Leishmania, induced by stress and differentiation that differs significantly from metazoa.

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Parasitic Adaptive Mechanisms in Infection by Leishmania

Cunningham

2002

Experimental and Molecular Pathology

211

122

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Down-regulation of MARCKS-related Protein (MRP) in Macrophages Infected with Leishmania

Cited by 37 publications

References 46 publications

MARCKS-related Protein (MRP) Is a Substrate for the Leishmania major Surface Protease Leishmanolysin (gp63)

MARCKS-related Protein (MRP) Is a Substrate for the Leishmania major Surface Protease Leishmanolysin (gp63)

Cell death in Leishmania induced by stress and differentiation: programmed cell death or necrosis?

Parasitic Adaptive Mechanisms in Infection by Leishmania

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