Expression of chicken gizzard RLC complements the cytokinesis and developmental defects of Dictyostelium RLC null cells.
Dictyostelium RLC null cells have defects in cytokinesis and development that can be rescued by expression of either the wild type Dictyostelium RLC or an RLC mutant that cannot be phosphorylated by MLCK (S13A) (Ostrow et al., 1994). The wild type and S13A mutant LCs rescued the cells equally well, despite the fact that RLC phosphorylation increases purified Dictyostelium myosin's activity 5-fold. In this report, we assess the ability of foreign RLCs to rescue the RLC null phenotype. The RLC from smooth muscle myosin, whose activity is tightly controlled by phosphorylation, rescued the null cell phenotype. The purified hybrid myosin had an activity and motility comparable to phosphorylated Dictyostelium myosin. In contrast, cells expressing skeletal muscle RLC were deficient in cytokinesis and development, despite having an activity and motility similar to that of myosin with the unposphorylatable S13A mutant RLC. Neither foreign LC was phosphorylated when expressed in Dictyostelium. These results suggest that the level of actin-activated ATPase activity and motility is not the sole determinant of proper myosin function in vivo. Other heavy chain/light chain interactions, which occur only with the native RLC and smooth muscle RLC, appear to be necessary for optimal function.
Chen
P
P
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA.
Chaudoir
B M
BM
Trybus
K M
KM
Chisholm
R L
RL
D000595:Amino Acid Sequence; D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D002645:Chickens; D004023:Dictyostelium / Q000378:metabolism*; D005895:Gizzard, Avian / Q000378:metabolism*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000378:metabolism; D009130:Muscle, Smooth / Q000378:metabolism; D018994:Myosin Light Chains / Q000096:biosynthesis / Q000502:physiology*; D010766:Phosphorylation; D016415:Sequence Alignment
D018994:Myosin Light Chains