TY - JOUR
T1 - RAC1-Dependent ORAI1 Translocation to the Leading Edge Supports Lamellipodia Formation and Directional Persistence
AU - Lopez-Guerrero, Aida M.
AU - Espinosa-Bermejo, Noelia
AU - Sanchez-Lopez, Irene
AU - Macartney, Thomas
AU - Pascual-Caro, Carlos
AU - Orantos-Aguilera, Yolanda
AU - Rodriguez-Ruiz, Lola
AU - Perez-Oliva, Ana B.
AU - Mulero, Victoriano
AU - Pozo-Guisado, Eulalia
AU - Martin-Romero, Francisco Javier
N1 - This work was supported by grants from the Spanish Ministerio de Ciencia, Innovación y Universidades (grants BFU2017-82716-P to FJMR, and BIO2017-84702-R to VM), and Junta de Extremadura (grants GR18084 and IB16088 to FJMR). All grants were co-financed by FEDER funds. ALG, NEB, and CPC were supported by predoctoral fellowships from the Spanish Ministerio de Economia y Competitividad (BES-2012-052061), Junta de Extremadura (PD18040), and Ministerio de Educación, Cultura y Deporte (FPU13/03430), respectively.
PY - 2020/4/20
Y1 - 2020/4/20
N2 - Tumor invasion requires efficient cell migration, which is achieved by the generation of persistent and polarized lamellipodia. The generation of lamellipodia is supported by actin dynamics at the leading edge where a complex of proteins known as the WAVE regulatory complex (WRC) promotes the required assembly of actin filaments to push the front of the cell ahead. By using an U2OS osteosarcoma cell line with high metastatic potential, proven by a xenotransplant in zebrafish larvae, we have studied the role of the plasma membrane Ca2+ channel ORAI1 in this process. We have found that epidermal growth factor (EGF) triggered an enrichment of ORAI1 at the leading edge, where colocalized with cortactin (CTTN) and other members of the WRC, such as CYFIP1 and ARP2/3. ORAI1-CTTN co-precipitation was sensitive to the inhibition of the small GTPase RAC1, an upstream activator of the WRC. RAC1 potentiated ORAI1 translocation to the leading edge, increasing the availability of surface ORAI1 and increasing the plasma membrane ruffling. The role of ORAI1 at the leading edge was studied in genetically engineered U2OS cells lacking ORAI1 expression that helped us to prove the key role of this Ca2+ channel on lamellipodia formation, lamellipodial persistence, and cell directness, which are required for tumor cell invasiveness in vivo.
AB - Tumor invasion requires efficient cell migration, which is achieved by the generation of persistent and polarized lamellipodia. The generation of lamellipodia is supported by actin dynamics at the leading edge where a complex of proteins known as the WAVE regulatory complex (WRC) promotes the required assembly of actin filaments to push the front of the cell ahead. By using an U2OS osteosarcoma cell line with high metastatic potential, proven by a xenotransplant in zebrafish larvae, we have studied the role of the plasma membrane Ca2+ channel ORAI1 in this process. We have found that epidermal growth factor (EGF) triggered an enrichment of ORAI1 at the leading edge, where colocalized with cortactin (CTTN) and other members of the WRC, such as CYFIP1 and ARP2/3. ORAI1-CTTN co-precipitation was sensitive to the inhibition of the small GTPase RAC1, an upstream activator of the WRC. RAC1 potentiated ORAI1 translocation to the leading edge, increasing the availability of surface ORAI1 and increasing the plasma membrane ruffling. The role of ORAI1 at the leading edge was studied in genetically engineered U2OS cells lacking ORAI1 expression that helped us to prove the key role of this Ca2+ channel on lamellipodia formation, lamellipodial persistence, and cell directness, which are required for tumor cell invasiveness in vivo.
KW - Calcium signalling
KW - Lamellipodia
UR - http://www.scopus.com/inward/record.url?scp=85083807969&partnerID=8YFLogxK
U2 - 10.1038/s41598-020-63353-5
DO - 10.1038/s41598-020-63353-5
M3 - Article
C2 - 32313105
SN - 2045-2322
VL - 10
SP - 1
EP - 18
JO - Scientific Reports
JF - Scientific Reports
M1 - 6580
ER -