Truncation of a mammalian myosin I results in loss of Ca2+-sensitive motility.
MYR-1, a mammalian class I myosin, consisting of a heavy chain and 4-6 associated calmodulins, is represented by the 130-kDa myosin I (or MI(130)) from rat liver. MI(130) translocates actin filaments in vitro in a Ca(2+)-regulated manner. A decrease in motility observed at higher Ca(2+) concentrations has been attributed to calmodulin dissociation. To investigate mammalian myosin I regulation, we have coexpressed in baculovirus calmodulin and an epitope-tagged 85-kDa fragment representing the amino-terminal catalytic "motor" domain and the first calmodulin-binding IQ domain of rat myr-1; we refer to this truncated molecule here as MI(1IQ). Association of calmodulin to MI(1IQ) is Ca(2+)-insensitive. MI(1IQ) translocates actin filaments in vitro at a rate resembling MI(130), but unlike MI(130), does not exhibit sensitivity to 0.1-100 micrometer Ca(2+). In addition to demonstrating successful expression of a functional truncated mammalian myosin I in vitro, these results indicate that: 1) Ca(2+)-induced calmodulin dissociation from MI(130) in the presence of actin is not from the first IQ domain, 2) velocity is not affected by the length of the IQ region, and 3) the Ca(2+) sensitivity of actin translocation exhibited by MI(130) involves 1 or more of the other 5 IQ domains and/or the carboxyl tail.
Perreault-Micale
C
C
Boston Biomedical Research Institute, Watertown, Massachusetts 02472, USA.
Shushan
A D
AD
Coluccio
L M
LM
D000199:Actins / Q000378:metabolism*; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002147:Calmodulin / Q000378:metabolism*; D017931:DNA Primers; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D024461:Myosin Type I; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism* / Q000502:physiology* / Q000648:ultrastructure; D016366:Open Reading Frames; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D017384:Sequence Deletion
D000199:Actins; D002147:Calmodulin; D017931:DNA Primers; C104297:Myo1b protein, rat; D011994:Recombinant Proteins; D024461:Myosin Type I; D009218:Myosins; D002118:Calcium