TY - JOUR
T1 - Cytoplasmic retention sites in p190RhoGEF confer anti-apoptotic activity to an EGFP-tagged protein
AU - Wu, Junhua
AU - Zhai, Jinbin
AU - Lin, Hong
AU - Nie, Zhenying
AU - Ge, Wei Wen
AU - García-Bermejo, Laura
AU - Muschel, Ruth J.
AU - Schlaepfer, William W.
AU - Cañete-Soler, Rafaela
N1 - Funding Information:
We thank Dr. Ben Margolis for providing the JIP-1 expression vector and JIP-1 antibody (JIP176), Dr. Wouter Moolenaar for providing the full-length p190RhoGEF cDNA and Dr. Randy Pittman for PC12 cells. The study was supported by NIH grant NS15722.
PY - 2003/9/10
Y1 - 2003/9/10
N2 - p190RhoGEF is a large multi-functional protein with guanine nucleotide exchange (GEF) activity. The C-terminal region of p190RhoGEF is a highly interactive domain that binds multiple factors, including proteins with anti-apoptotic activities. We now report that transfection of EGFP-tagged p190RhoGEF protects Neuro 2a cells from stress-induced apoptosis and that anti-apoptotic activity is localized to cytoplasmic retention sequences (CRS-1 and CRS-2) in the C-terminal region of p190RhoGEF. Cytoplasmic retention is conferred to an EGFP fluorescent marker when fused to either CRS-1 or CRS-2. Both cytoplasmic retention and anti-apoptotic activity are lost by deleting CRS-1 and CRS-2 in the p190RhoGEF sequence and can be recovered by restoring either CRS-1 or CRS-2 to the EGFP-tagged sequence. Since the CRS-1 and CRS-2 contain the JIP-1 and 14-3-3 binding sites, we propose that anti-apoptotic activity may be conferred by the binding of p190RhoGEF to JIP-1 or 14-3-3, possibly by altering their interactive properties or nucleocytoplasmic movements. Taken together, our findings support a model whereby multiple interactions of p190RhoGEF confer homeostatic properties to differentiated neurons and may link neuronal homeostasis to the regulation of NF-L expression.
AB - p190RhoGEF is a large multi-functional protein with guanine nucleotide exchange (GEF) activity. The C-terminal region of p190RhoGEF is a highly interactive domain that binds multiple factors, including proteins with anti-apoptotic activities. We now report that transfection of EGFP-tagged p190RhoGEF protects Neuro 2a cells from stress-induced apoptosis and that anti-apoptotic activity is localized to cytoplasmic retention sequences (CRS-1 and CRS-2) in the C-terminal region of p190RhoGEF. Cytoplasmic retention is conferred to an EGFP fluorescent marker when fused to either CRS-1 or CRS-2. Both cytoplasmic retention and anti-apoptotic activity are lost by deleting CRS-1 and CRS-2 in the p190RhoGEF sequence and can be recovered by restoring either CRS-1 or CRS-2 to the EGFP-tagged sequence. Since the CRS-1 and CRS-2 contain the JIP-1 and 14-3-3 binding sites, we propose that anti-apoptotic activity may be conferred by the binding of p190RhoGEF to JIP-1 or 14-3-3, possibly by altering their interactive properties or nucleocytoplasmic movements. Taken together, our findings support a model whereby multiple interactions of p190RhoGEF confer homeostatic properties to differentiated neurons and may link neuronal homeostasis to the regulation of NF-L expression.
KW - Anti-apoptosis
KW - Cytoplasmic retention
KW - Neuronal homeostasis
KW - Nucleocytoplasmic movements
KW - p190RhoGEF
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UR - http://www.scopus.com/inward/citedby.url?scp=0141671835&partnerID=8YFLogxK
U2 - 10.1016/S0169-328X(03)00263-8
DO - 10.1016/S0169-328X(03)00263-8
M3 - Article
C2 - 14499478
AN - SCOPUS:0141671835
SN - 0169-328X
VL - 117
SP - 27
EP - 38
JO - Molecular Brain Research
JF - Molecular Brain Research
IS - 1
ER -