Phosphoinositide-specific phospholipase C in oat roots: association with the actin cytoskeleton.
Phosphoinositide-specific phospholipase C (PI-PLC) activities are involved in mediating plant cell responses to environmental stimuli. Two variants of PI-PLC have been partially purified from the roots of oat seedlings; one cytosolic and one particulate. Although the cytosolic enzyme was significantly purified, the activity still co-migrated with a number of other proteins on heparin HPLC and also on size-exclusion chromatography. The partially purified PI-PLC was tested by Western blotting, and we found that actin and actin-binding proteins, profilin and tropomyosin, co-purified with cytosolic phospholipase C. After a non-ionic detergent (Triton X-100) treatment, PI-PLC activities still remained with the actin cytoskeleton. The effects of phalloidin and F-buffer confirmed this association; these conditions, which favor actin polymerization, decreased the release of PI-PLC from the cytoskeleton. The treatments of latrunculin and G-buffer, the conditions that favor actin depolymerization, increased the release of PI-PLC from the cytoskeleton. These results suggest that oat PI-PLC associates with the actin cytoskeleton.
Huang
Chiung-Hua
CH
Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Science and Technology, Pei-tun District, Taichung 40601, Taiwan. chhuang@ctust.edu.tw
Crain
Richard C
RC
D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D000818:Animals; D018554:Avena / Q000187:drug effects / Q000201:enzymology*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002417:Cattle; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D003429:Cross Reactions / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects / Q000201:enzymology*; D003902:Detergents / Q000494:pharmacology; D007106:Immune Sera / Q000494:pharmacology; D047468:Immunoprecipitation; D010590:Phalloidine / Q000378:metabolism; D054801:Phosphoinositide Phospholipase C / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D054799:Phospholipase C beta / Q000378:metabolism; D018517:Plant Roots / Q000187:drug effects / Q000201:enzymology*; D051304:Profilins / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D012995:Solubility / Q000187:drug effects; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D053778:Thiazolidines / Q000494:pharmacology
D000199:Actins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D003902:Detergents; D007106:Immune Sera; D051304:Profilins; D053778:Thiazolidines; D010590:Phalloidine; D054801:Phosphoinositide Phospholipase C; D054799:Phospholipase C beta; C037068:latrunculin B