A critical period requiring Rho proteins for cell cycle progression uncovered by reversible protein transduction in endothelial cells.
The time-specific requirement of Rho proteins for the S phase progression of vascular endothelial cells was determined by reversibly introducing inhibitor proteins with a cell-penetrating peptide. We found evidence of the reversibility of protein transduction. The removal of extracellular protein caused the transduced protein to decay in a manner sensitive to low temperatures. The time required for a 50% decay correlated with the protein size. The time-specific transduction of the inhibitor proteins uncovered a critical period requiring Rho proteins in the G1-S transition phase. Reversible protein transduction may thus be a powerful tool to investigate the time-specific role of signaling proteins.
Hirano
Katsuya
K
Division of Molecular Cardiology, Research Institute of Angiocardiology, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. khirano@molcar.med.kyushu-u.ac.jp
Hirano
Mayumi
M
Nishimura
Junji
J
Kanaide
Hideo
H
D000818:Animals; D001011:Aorta / Q000166:cytology; D002417:Cattle; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology*; D004730:Endothelium, Vascular / Q000378:metabolism; D005434:Flow Cytometry; D016193:G1 Phase; D049452:Green Fluorescent Proteins; D015151:Immunoblotting; D008164:Luminescent Proteins / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000378:metabolism; D016196:S Phase; D015398:Signal Transduction; D013696:Temperature; D013997:Time Factors; D020741:rho GTP-Binding Proteins / Q000378:metabolism*
D008164:Luminescent Proteins; D011994:Recombinant Proteins; D016160:Retinoblastoma Protein; D049452:Green Fluorescent Proteins; D020741:rho GTP-Binding Proteins