The roles of Cdc42 and Rac1 in the formation of plasma membrane protrusions in cancer epithelial HeLa cells.
Cell Membrane
/ metabolism
Epithelial Cells
/ metabolism
Female
Guanine Nucleotide Exchange Factors
/ genetics
HeLa Cells
Humans
Pseudopodia
/ metabolism
Signal Transduction
/ genetics
Transfection
Uterine Cervical Neoplasms
/ metabolism
cdc42 GTP-Binding Protein
/ genetics
rac1 GTP-Binding Protein
/ genetics
Cdc42
DOCK10
DOCK9
Filopodia
Membrane blebs
Membrane ruffles
Rac1
Rho GTPase
Journal
Molecular biology reports
ISSN: 1573-4978
Titre abrégé: Mol Biol Rep
Pays: Netherlands
ID NLM: 0403234
Informations de publication
Date de publication:
May 2021
May 2021
Historique:
received:
22
01
2021
accepted:
27
05
2021
pubmed:
11
6
2021
medline:
31
12
2021
entrez:
10
6
2021
Statut:
ppublish
Résumé
The inducible model of clones generated from the cervical cancer epithelial HeLa cell line has shown the role of DOCK10 as a guanine-nucleotide exchange factor for Rho GTPases Cdc42 and Rac1 and as an inducer of filopodia and plasma membrane (PM) ruffles. In this model, constitutively active (CA) mutants of Cdc42 and Rac1 promote filopodia and ruffles, respectively, as in other models. DOCK9 also induces filopodia and ruffles, although ruffling activity is less prominent. By exploiting this model further, the aim of this work is to provide a more complete understanding of the role of Cdc42 and Rac1, and their interactions with DOCK10 and DOCK9, in regulation of PM protrusions. New clones have been generated from HeLa, including single clones expressing one form of wild-type (WT) or dominant negative (DN) Cdc42 or Rac1, and double clones co-expressing one of them together with either DOCK10 or DOCK9. Expression of WT- and DN-Cdc42 induced filopodia. WT-Cdc42 and, especially, DN-Cdc42 also gave rise to veil protrusions, which were neutralized by DOCK10. Moreover, DN-Cdc42 stimulated the emergence of ruffles, further increased by DOCK10, and WT-Cdc42 also augmented ruffles in presence of DOCK9 and DOCK10. WT-Rac1 greatly increased PM blebbing, as did DN-Rac1 more moderately. In both cases, blebs were enhanced by DOCK10. DN-Rac1 and CA-Rac1 moderately raised filopodia, and DOCK10 and DOCK9 had opposed effects on filopodia (up and down, respectively) in presence of WT-Rac1. As conclusions, we highlight that Cdc42 promotes filopodia regardless of its conformational state, and Rac1 induces blebs in conformations other than CA, especially WT-Rac1, in the inducible HeLa clones. The model could be useful to learn more about the mechanisms underlying PM protrusions.
Identifiants
pubmed: 34110575
doi: 10.1007/s11033-021-06443-5
pii: 10.1007/s11033-021-06443-5
doi:
Substances chimiques
DOCK9 protein, human
0
Dock10 protein, human
0
Guanine Nucleotide Exchange Factors
0
RAC1 protein, human
0
CDC42 protein, human
EC 3.6.5.2
cdc42 GTP-Binding Protein
EC 3.6.5.2
rac1 GTP-Binding Protein
EC 3.6.5.2
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4285-4294Subventions
Organisme : Instituto de Salud Carlos III
ID : PI10/01226
Organisme : Fundación Séneca
ID : 08721/PI/08
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