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Identification of Giardia lamblia DHHC proteins and the role of protein S-palmitoylation in the encystation process.

Merino MC, Zamponi N, Vranych CV, Touz MC, Rópolo AS - PLoS Negl Trop Dis (2014)

Bottom Line: With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome.These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades.Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigación Médica Mercedes y Martín Ferreyra, INIMEC - Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Córdoba, Córdoba, Argentina.

ABSTRACT
Protein S-palmitoylation, a hydrophobic post-translational modification, is performed by protein acyltransferases that have a common DHHC Cys-rich domain (DHHC proteins), and provides a regulatory switch for protein membrane association. In this work, we analyzed the presence of DHHC proteins in the protozoa parasite Giardia lamblia and the function of the reversible S-palmitoylation of proteins during parasite differentiation into cyst. Two specific events were observed: encysting cells displayed a larger amount of palmitoylated proteins, and parasites treated with palmitoylation inhibitors produced a reduced number of mature cysts. With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome. These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades. Although all Giardia DHHC proteins were found to be present in trophozoites and encysting cells, these proteins showed a different intracellular localization in trophozoites and seemed to be differently involved in the encystation process when they were overexpressed. dhhc transgenic parasites showed a different pattern of cyst wall protein expression and yielded different amounts of mature cysts when they were induced to encyst. Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.

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Analysis of palmitoylated proteins in dhhc transgenic growing and encysting parasites displays a similar pattern to wild type parasites.Giardia trophozoites (T) or encysting trophozoites (ET) were labeled with [3H]-palmitic acid and loaded onto SDS-PAGE. Samples were then analyzed by autoradiography. The approximate sizes are indicated on the right in kDa.
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pntd-0002997-g010: Analysis of palmitoylated proteins in dhhc transgenic growing and encysting parasites displays a similar pattern to wild type parasites.Giardia trophozoites (T) or encysting trophozoites (ET) were labeled with [3H]-palmitic acid and loaded onto SDS-PAGE. Samples were then analyzed by autoradiography. The approximate sizes are indicated on the right in kDa.

Mentions: The different localization of DHHC-HA proteins in trophozoites and the differential effect of DHHC overexpression in encystation prompted us to evaluate the palmitoylation pattern in the dhhc transgenic parasites (Figure 10). gla_1908, gla_2116, gla_16928, and gla_8711 transgenic trophozoites or encysting parasites displayed a similar global protein palmitoylation pattern compared to wild type (Figure 1A). Mass spectrometry-based proteomics analyses will be necessary to accurately identify any differences in the palmitoylation substrates among the dhhc transgenic parasites.


Identification of Giardia lamblia DHHC proteins and the role of protein S-palmitoylation in the encystation process.

Merino MC, Zamponi N, Vranych CV, Touz MC, Rópolo AS - PLoS Negl Trop Dis (2014)

Analysis of palmitoylated proteins in dhhc transgenic growing and encysting parasites displays a similar pattern to wild type parasites.Giardia trophozoites (T) or encysting trophozoites (ET) were labeled with [3H]-palmitic acid and loaded onto SDS-PAGE. Samples were then analyzed by autoradiography. The approximate sizes are indicated on the right in kDa.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109852&req=5

pntd-0002997-g010: Analysis of palmitoylated proteins in dhhc transgenic growing and encysting parasites displays a similar pattern to wild type parasites.Giardia trophozoites (T) or encysting trophozoites (ET) were labeled with [3H]-palmitic acid and loaded onto SDS-PAGE. Samples were then analyzed by autoradiography. The approximate sizes are indicated on the right in kDa.
Mentions: The different localization of DHHC-HA proteins in trophozoites and the differential effect of DHHC overexpression in encystation prompted us to evaluate the palmitoylation pattern in the dhhc transgenic parasites (Figure 10). gla_1908, gla_2116, gla_16928, and gla_8711 transgenic trophozoites or encysting parasites displayed a similar global protein palmitoylation pattern compared to wild type (Figure 1A). Mass spectrometry-based proteomics analyses will be necessary to accurately identify any differences in the palmitoylation substrates among the dhhc transgenic parasites.

Bottom Line: With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome.These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades.Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.

View Article: PubMed Central - PubMed

Affiliation: Instituto de Investigación Médica Mercedes y Martín Ferreyra, INIMEC - Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Nacional de Córdoba, Córdoba, Argentina.

ABSTRACT
Protein S-palmitoylation, a hydrophobic post-translational modification, is performed by protein acyltransferases that have a common DHHC Cys-rich domain (DHHC proteins), and provides a regulatory switch for protein membrane association. In this work, we analyzed the presence of DHHC proteins in the protozoa parasite Giardia lamblia and the function of the reversible S-palmitoylation of proteins during parasite differentiation into cyst. Two specific events were observed: encysting cells displayed a larger amount of palmitoylated proteins, and parasites treated with palmitoylation inhibitors produced a reduced number of mature cysts. With bioinformatics tools, we found nine DHHC proteins, potential protein acyltransferases, in the Giardia proteome. These proteins displayed a conserved structure when compared to different organisms and are distributed in different monophyletic clades. Although all Giardia DHHC proteins were found to be present in trophozoites and encysting cells, these proteins showed a different intracellular localization in trophozoites and seemed to be differently involved in the encystation process when they were overexpressed. dhhc transgenic parasites showed a different pattern of cyst wall protein expression and yielded different amounts of mature cysts when they were induced to encyst. Our findings disclosed some important issues regarding the role of DHHC proteins and palmitoylation during Giardia encystation.

Show MeSH
Related in: MedlinePlus