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Barboza, B. R.

Publications and source records attributed to Barboza, B. R..

3 recordsLinked to original sources

ST8Sia2 polysialyltransferase protects against infection by Trypanosoma cruzi

Glycosylation is one of the most structurally and functionally diverse co- and post-translational modifications in a cell. Addition and removal of glycans, especially to proteins and lipids, characterize this process which have important implications in several biological processes. In mammals, the repeated enzymatic addition of a sialic acid unit to underlying sialic acids (Sia) by polysialyltransferases, including ST8Sia2, leads to the formation of a sugar polymer called polysialic acid (polySia). The functional relevance of polySia has been extensively demonstrated in the nervous system. However, the role of polysialylation in infection is still poorly explored. Previous reports have shown that Trypanosoma cruzi (T. cruzi), a flagellated parasite that causes Chagas disease (CD), changes host sialylation of glycoproteins. To understand the role of host polySia during T. cruzi infection, we used a combination of in silico and experimental tools. We observed that T. cruzi reduces both the expression of the ST8Sia2 and the polysialylation of target substrates. We also found that chemical and genetic inhibition of host ST8Sia2 increased the parasite load in mammalian cells. These findings suggest a novel approach to interfere with parasite infections through modulation of host polysialylation. AUTHOR SUMMARYGlycosylation is a co- and/or post-translational modification regulated by the addition and removal of glycans. This process shapes the cellular glycome, which in turn, holds significant implications in various biological processes. Trypanosoma cruzi (T. cruzi), the etiological agent of Chagas disease, a globally concerning neglected tropical disease affecting 6 to 8 million individuals worldwide, exerts a profound influence on host glycoprotein sialylation. Remarkably, T. cruzi is incapable of synthesizing sialic acid (Sia) and relies on acquiring it from host glycoconjugates. In mammals, the formation of polysialic acid (polySia) is mediated by polysialyltransferases, such as ST8Sia2. The functional relevance of polySia has been extensively documented in the nervous system. Nevertheless, its role within the context of infectious processes remains largely unexplored. Herein, we demonstrate that in T. cruzi-infected host cells, the expression of the ST8Sia2 enzyme is downregulated, resulting in diminished levels of polysialylation. Furthermore, a reduction in the levels of NCAM1 and SCN5A was observed, which can be attributed to the decreased host polysialylation. Moreover, enzymatic removal of polySia, along with chemical inhibition and genetic silencing of ST8Sia2, led to a marked increase in the number of intracellular parasites. We posit that ST8Sia2 inhibition favors T. cruzi infection, thereby elucidating novel avenues for understanding the mechanisms associated with Chagas disease pathogenesis, prominently featuring the pivotal role of host polysialylation.

microbiology↗

ArtinM Cytotoxicity in B cells derived from Non-Hodgkins Lymphoma is regulated by CD45 phosphatase activity and Src family kinases

Receptors on the immune cell surface have a variety of glycans that may account for the immunomodulation induced by lectins, which have a carbohydrate recognition domain (CRD) that binds to monosaccharides or oligosaccharides in a specific manner. ArtinM, a D-mannose-binding lectin obtained from Artocarpus heterophyllus, has affinity for the N-glycans core. Immunomodulation by ArtinM toward the Th1 phenotype occurs via its interaction with TLR2/CD14 N-glycans on antigen-presenting cells, as well as recognition of CD3{gamma} N-glycans on murine CD4+ and CD8+ T cells. ArtinM exerts a cytotoxic effect on Jurkat human leukemic T cell line and human myeloid leukemia cell line (NB4). The current study evaluated the effects of ArtinM on murine and human B cells derived from non-Hodgkins lymphoma. We found that murine B cells are recognized by ArtinM via the CRD, and the ArtinM stimulus did not augment the proliferation rate or production of IL-2. However, murine B cells incubation with ArtinM augmented the rate of apoptosis, and this cytotoxic effect of ArtinM was also seen in human B cell lines sourced from non-Hodgkins lymphoma Raji cell line. This cytotoxic effect was inhibited by the phosphatase activity of CD45 on Lck, and the protein kinases of the Src family contribute to cell death triggered by ArtinM.

cell biology↗

Protein arginylation is regulated during SARS-CoV-2 infection

In 2019, the world witnessed the onset of an unprecedented pandemic. In September 2021, the infection by SARS-CoV-2 had already been responsible for the death of more than 4 million people worldwide. Recently, we and other groups discovered that SARS-CoV-2 infection induces ER-stress and activation of unfolded protein response (UPR) pathway. The degradation of misfolded/unfolded proteins is an essential element of proteostasis and occurs mainly in lysosomes or proteasomes. The N-terminal arginylation of proteins is characterized as an inducer of ubiquitination and proteasomal degradation by the N-end rule pathway. Here we present, for the first time, data on the role of arginylation during SARS-CoV-2 infection. We studied the modulation of protein arginylation in Vero CCL-81 and Calu-3 cells infected after 2h, 6h, 12h, 24h, and 48h. A reanalysis of in vivo and in vitro public omics data combined with immunoblotting was performed to measure the levels of ATE1 and arginylated proteins. This regulation is seen specifically during infections by coronaviruses. We demonstrate that during SARS-CoV-2 infection there is an increase in the expression of the ATE1 enzyme associated with regulated levels of specific arginylated proteins. On the other hand, infected macrophages showed no ATE1 regulation. An important finding revealed that modulation of the N-end rule pathway differs between different types of infected cells. We also confirmed the potential of tannic acid to reduce viral load, and furthermore, to modulate ATE1 levels during infection. In addition, the arginylation inhibitor merbromin (MER) is also capable of both reducing viral load and reducing ATE1 levels. Taken together, these data show the importance of arginylation during the progression of SARS-CoV-2 infection and open the door for future studies that may unravel the role of ATE1 and its inhibitors in pathogen infection.

cell biology↗