2',3'-Cyclic nucleotide 3'-phosphodiesterase binds to actin-based cytoskeletal elements in an isoprenylation-independent manner.
2',3'-Cyclic nucleotide 3'-phosphodiesterase (CNP) is an isoprenylated protein enriched in myelin and oligodendrocytes but also present in several other tissues at low levels. CNP binds avidly to membranes and in addition possesses several characteristics of cytoskeletal proteins. The role of isoprenylation in the association of CNP with the cytoskeleton was analyzed by ectopic expression in L cells of epitope-tagged CNP1 and a non-isoprenylated mutant CNP1. Using nonionic detergent extraction, drug-mediated cytoskeletal disruption, and coimmunoprecipitation with an anti-actin antibody, we show that CNP1 is associated with actin-based cytoskeletal elements independently of its isoprenylation status. A control protein, p21c-H-ras, which is also modified by isoprenylation at its carboxyl-terminus, does not bind to cytoskeletal structures as judged by the same criteria. We present a model that accounts for the association of CNP1 with membranes and the cytoskeleton.
De Angelis
D A
DA
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Braun
P E
PE
D063309:2',3'-Cyclic Nucleotide 3'-Phosphodiesterase; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000302:isolation & purification / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D002458:Cell Fractionation; D003078:Colchicine / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D003902:Detergents; D019085:Fluorescent Antibody Technique, Direct; D007739:L Cells / Q000737:chemistry / Q000201:enzymology; D051379:Mice; D010727:Phosphoric Diester Hydrolases; D011233:Precipitin Tests; D011485:Protein Binding / Q000502:physiology; D017368:Protein Prenylation / Q000502:physiology*; D012995:Solubility; D014162:Transfection
D000199:Actins; D003902:Detergents; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D010727:Phosphoric Diester Hydrolases; D063309:2',3'-Cyclic Nucleotide 3'-Phosphodiesterase; C571338:Cnp protein, mouse; D003078:Colchicine