Role of myosin II tail sequences in its function and localization at the cleavage furrow in Dictyostelium.
Cytoplasmic myosin II accumulates in the cleavage furrow and provides the force for cytokinesis in animal and amoeboid cells. One model proposes that a specific domain in the myosin II tail is responsible for its localization, possibly by interacting with a factor concentrated in the equatorial region. To test this possibility, we have expressed myosins carrying mutations in the tail domain in a strain of Dictyostelium cells from which the endogenous myosin heavy chain gene has been deleted. The mutations used in this study include four internal tail deletions: Mydelta824-941, Mydelta943-1464, Mydelta943-1194 and Mydelta1156-1464. Contrary to the prediction of the hypothesis, immunofluorescence staining demonstrated that all mutant myosins were able to move toward the furrow region. Chimeric myosins, which consisted of a Dictyostelium myosin head and chicken skeletal myosin tail, also efficiently localized to the cleavage furrow. All these deletion and chimeric mutant myosins, except for Mydelta943-1464, the largest deletion mutant, were able to support cytokinesis in suspension. Our data suggest that there is no single specific domain in the tail of Dictyostelium myosin II that is required for its functioning at and localization to the cleavage furrow.
Shu
S
S
Biomolecular Research Group, National Institute for Advanced Interdisciplinary Research, Tsukuba, Ibaraki 305-8562, Japan.
Lee
R J
RJ
LeBlanc-Straceski
J M
JM
Uyeda
T Q
TQ
D000818:Animals; D001483:Base Sequence; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D004023:Dictyostelium / Q000166:cytology / Q000235:genetics* / Q000502:physiology*; D017125:Genes, Protozoan; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009218:Myosins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000502:physiology; D017384:Sequence Deletion; D013347:Subcellular Fractions / Q000502:physiology
D017931:DNA Primers; D011993:Recombinant Fusion Proteins; D009218:Myosins