TY - JOUR
T1 - Giardia intestinalis coiled-coil cytolinker protein 259 interacts with actin and tubulin
AU - Rojas-Gutiérrez, Omar
AU - Pérez-Rangel, Armando
AU - Castillo-Romero, Araceli
AU - Tapia-Ramírez, José
AU - Ríos-Castro, Emmanuel
AU - Camacho-Nuez, Minerva
AU - Giono-Cerezo, Silvia
AU - Nogueda-Torres, Benjamín
AU - León-Avila, Gloria
AU - Hernández, José Manuel
N1 - Publisher Copyright:
© 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.
PY - 2021/3
Y1 - 2021/3
N2 - 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.
AB - 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.
KW - Coiled-coil
KW - Cytoskeleton
KW - Giardia intestinalis
KW - Vesicular traffic
UR - http://www.scopus.com/inward/record.url?scp=85099926408&partnerID=8YFLogxK
U2 - 10.1007/s00436-021-07062-6
DO - 10.1007/s00436-021-07062-6
M3 - Artículo
C2 - 33515065
AN - SCOPUS:85099926408
SN - 0932-0113
VL - 120
SP - 1067
EP - 1076
JO - Parasitology Research
JF - Parasitology Research
IS - 3
ER -