Regulation of cytoplasmic dynein ATPase by Lis1.
Mutations in Lis1 cause classical lissencephaly, a developmental brain abnormality characterized by defects in neuronal positioning. Over the last decade, a clear link has been forged between Lis1 and the microtubule motor cytoplasmic dynein. Substantial evidence indicates that Lis1 functions in a highly conserved pathway with dynein to regulate neuronal migration and other motile events. Yeast two-hybrid studies predict that Lis1 binds directly to dynein heavy chains (Sasaki et al., 2000; Tai et al., 2002), but the mechanistic significance of this interaction is not well understood. We now report that recombinant Lis1 binds to native brain dynein and significantly increases the microtubule-stimulated enzymatic activity of dynein in vitro. Lis1 does this without increasing the proportion of dynein that binds to microtubules, indicating that Lis1 influences enzymatic activity rather than microtubule association. Dynein stimulation in vitro is not a generic feature of microtubule-associated proteins, because tau did not stimulate dynein. To our knowledge, this is the first indication that Lis1 or any other factor directly modulates the enzymatic activity of cytoplasmic dynein. Lis1 must be able to homodimerize to stimulate dynein, because a C-terminal fragment (containing the dynein interaction site but missing the self-association domain) was unable to stimulate dynein. Binding and colocalization studies indicate that Lis1 does not interact with all dynein complexes found in the brain. We propose a model in which Lis1 stimulates the activity of a subset of motors, which could be particularly important during neuronal migration and long-distance axonal transport.
Department of Biological Sciences, University of South Carolina, Columbia, South Carolina 29208, USA.
Mariano T
MT
Mesngon
Cataldo
C
Tarricone
Sachin
S
Hebbar
Aimee M
AM
Guillotte
E William
EW
Schmitt
Lorene
L
Lanier
Andrea
A
Musacchio
Stephen J
SJ
King
Deanna S
DS
Smith
D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; D017398:Alternative Splicing; D000818:Animals; D001370:Axonal Transport / Q000502:physiology; D001921:Brain / Q000502:physiology; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology; D004398:Dyneins / Q000378:metabolism*; D014644:Genetic Variation; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D018411:Spodoptera; D014162:Transfection
D008869:Microtubule-Associated Proteins; D011994:Recombinant Proteins; D043203:1-Alkyl-2-acetylglycerophosphocholine Esterase; C513190:Pafah1b1 protein, mouse; D004398:Dyneins