Slow axonal transport of soluble actin with actin depolymerizing factor, cofilin, and profilin suggests actin moves in an unassembled form.
We examined the axonal transport of actin and its monomer binding proteins, actin depolymerizing factor, cofilin, and profilin, in the chicken sciatic nerve following injection of [35S]methionine into the lumbar spinal cord. At intervals up to 20 days after injection, nerves were cut into 1-cm segments and separated into Triton X-100-soluble and particulate fractions. Actin and its binding proteins were then isolated by affinity chromatography on DNase I-Sepharose and by one- and two-dimensional polyacrylamide gel electrophoresis. Fluorographic analysis showed that the specific activity of soluble actin was two to three times that of its particulate form and that soluble actin, cofilin, actin depolymerizing factor, and profilin were transported at similar rates in slow component b of axonal flow. Our data strongly support the view that the mobile form of actin in slow transport is soluble and that a substantial amount of this actin may travel as a complex with actin depolymerizing factor, cofilin, and profilin. Along labeled nerves the specific activity of the unphosphorylated form of actin depolymerizing factor, which binds actin, was not significantly different from that of its "inactive" phosphorylated form. This constancy in specific activity suggests that continuous inactivation and reactivation of actin depolymerizing factor occur during transport, which could contribute to the exchange of soluble actin with the filamentous actin pool.
Mills
R G
RG
Neuroscience Centre, University of Otago Medical School, Dunedin, New Zealand.
Minamide
L S
LS
Yuan
A
A
Bamburg
J R
JR
Bray
J J
JJ
D051339:Actin Depolymerizing Factors; D000199:Actins / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001370:Axonal Transport / Q000502:physiology*; D002645:Chickens; D002846:Chromatography, Affinity; D003285:Contractile Proteins; D003851:Deoxyribonucleases; D051345:Destrin; D015180:Electrophoresis, Gel, Two-Dimensional; D008840:Microfilament Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000302:isolation & purification / Q000378:metabolism*; D009474:Neurons / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D010766:Phosphorylation; D051304:Profilins; D012584:Sciatic Nerve / Q000737:chemistry / Q000166:cytology / Q000502:physiology; D012685:Sepharose; D012995:Solubility
D051339:Actin Depolymerizing Factors; D000199:Actins; D003285:Contractile Proteins; D051345:Destrin; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D051304:Profilins; D012685:Sepharose; D003851:Deoxyribonucleases