Translocation and association of ROCK-II with RhoA and HSP27 during contraction of rabbit colon smooth muscle cells.
The focus of the paper is to understand the role of HSP27 in mediating the association of RhoA with ROCK-II in sustained contraction of smooth muscle cells from the rabbit colon. In circular smooth muscle cells; acetylcholine-induced contraction (10(-7)M) was associated with translocation of ROCK-II to the particulate fraction, which remained sustained at 4 min after stimulation (135.1+/-8.1% increase, P </= 0.05). There was also an increased association of ROCK-II with RhoA particulate fraction (147.46+/-9.31 and 148.22+/-9.41, n = 3, P </= 0.05) and with HSP27 (155.6+/-10.7% increase, P </= 0.05) in the particulate fraction. Pre-incubation of cells with Y27632 resulted in the inhibition of the association of ROCK-II with RhoA in the particulate fraction. Acetylcholine (10(-7)M) induced sustained phosphorylation of MLC (122.75+/-9.97%, P </= 0.05 and 174.65+/-28.36%, P </= 0.05 increase in the di phospho-MLC at 30s and 4 min, respectively), which was inhibited upon pre-incubation with Y27632. Results suggest that ROCK-II undergoes a translocation to the particulate fraction with RhoA and with HSP27, suggesting that translocation and association of ROCK-II with RhoA is mediated by HSP27. Maintenance of the functional association of RhoA with ROCK-II in the particulate fraction mediated by HSP27 appears to be important to retain MLC in the phosphorylated state and hence the sustained contraction.
Patil
Suresh B
SB
Department of Pediatrics, University of Michigan, Ann Arbor, MI 48109, USA.
Tsunoda
Yasuhiro
Y
Pawar
Mercy D
MD
Bitar
Khalil N
KN
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D015153:Blotting, Western; D003106:Colon / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000737:chemistry*; D047908:Intracellular Signaling Peptides and Proteins; D008954:Models, Biological; D009119:Muscle Contraction; D009130:Muscle, Smooth / Q000166:cytology*; D009363:Neoplasm Proteins / Q000737:chemistry*; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000378:metabolism; D021381:Protein Transport; D011725:Pyridines / Q000737:chemistry; D011817:Rabbits; D013997:Time Factors; D014162:Transfection; D014665:Vasodilator Agents / Q000737:chemistry; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein / Q000737:chemistry* / Q000378:metabolism
D006360:Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D011725:Pyridines; D014665:Vasodilator Agents; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; D000109:Acetylcholine