Giardia intestinalis coiled-coil cytolinker protein 259 interacts with actin and tubulin

Omar Rojas-Gutiérrez, Armando Pérez-Rangel, Araceli Castillo-Romero, José Tapia-Ramírez, Emmanuel Ríos-Castro, Minerva Camacho-Nuez, Silvia Giono-Cerezo, Benjamín Nogueda-Torres, Gloria León-Avila, José Manuel Hernández

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Giardia intestinalis is a human parasite that causes a diarrheal disease in developing countries. G. intestinalis has a cytoskeleton (CSK) composed of microtubules and microfilaments, and the Giardia genome does not code for the canonical CSK-binding proteins described in other eukaryotic cells. To identify candidate actin and tubulin cross-linking proteins, we performed a BLAST analysis of the Giardia genome using a spectraplakins consensus sequence as a query. Based on the highest BLAST score, we selected a 259-kDa sequence designated as a cytoskeleton linker protein (CLP259). The sequence was cloned in three fragments and characterized by immunoprecipitation, confocal microscopy, and mass spectrometry (MS). CLP259 was located in the cytoplasm in the form of clusters of thick rods and colocalized with actin at numerous sites and with tubulin in the median body. Immunoprecipitation followed by mass spectrometry revealed that CLP259 interacts with structural proteins such as giardins, SALP-1, axonemal, and eight coiled-coils. The vesicular traffic proteins detected were Mu adaptin, Vacuolar ATP synthase subunit B, Bip, Sec61 alpha, NSF, AP complex subunit beta, and dynamin. These results indicate that CLP259 in trophozoites is a CSK linker protein for actin and tubulin and could act as a scaffold protein driving vesicular traffic.

Original languageEnglish
Pages (from-to)1067-1076
Number of pages10
JournalParasitology Research
Volume120
Issue number3
DOIs
StatePublished - Mar 2021
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.

Keywords

  • Coiled-coil
  • Cytoskeleton
  • Giardia intestinalis
  • Vesicular traffic

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