TY - JOUR
T1 - TRPM7 regulates myosin IIA filament stability and protein localization by heavy chain phosphorylation
AU - Clark, Kristopher
AU - Middelbeek, Jeroen
AU - Lasonder, Edwin
AU - Dulyaninova, Natalya G
AU - Morrice, Nick A
AU - Ryazanov, Alexey G
AU - Bresnick, Anne R
AU - Figdor, Carl G
AU - van Leeuwen, Frank N
PY - 2008/5/9
Y1 - 2008/5/9
N2 - Deregulation of myosin II-based contractility contributes to the pathogenesis of human diseases, such as cancer, which underscores the necessity for tight spatial and temporal control of myosin II activity. Recently, we demonstrated that activation of the mammalian alpha-kinase TRPM7 inhibits myosin II-based contractility in a Ca(2+)- and kinase-dependent manner. However, the molecular mechanism is poorly defined. Here, we demonstrate that TRPM7 phosphorylates the COOH-termini of both mouse and human myosin IIA heavy chains--the COOH-terminus being a region that is critical for filament stability. Phosphorylated residues were mapped to Thr1800, Ser1803 and Ser1808. Mutation of these residues to alanine and that to aspartic acid lead to an increase and a decrease, respectively, in myosin IIA incorporation into the actomyosin cytoskeleton and accordingly affect subcellular localization. In conclusion, our data demonstrate that TRPM7 regulates myosin IIA filament stability and localization by phosphorylating a short stretch of amino acids within the alpha-helical tail of the myosin IIA heavy chain.
AB - Deregulation of myosin II-based contractility contributes to the pathogenesis of human diseases, such as cancer, which underscores the necessity for tight spatial and temporal control of myosin II activity. Recently, we demonstrated that activation of the mammalian alpha-kinase TRPM7 inhibits myosin II-based contractility in a Ca(2+)- and kinase-dependent manner. However, the molecular mechanism is poorly defined. Here, we demonstrate that TRPM7 phosphorylates the COOH-termini of both mouse and human myosin IIA heavy chains--the COOH-terminus being a region that is critical for filament stability. Phosphorylated residues were mapped to Thr1800, Ser1803 and Ser1808. Mutation of these residues to alanine and that to aspartic acid lead to an increase and a decrease, respectively, in myosin IIA incorporation into the actomyosin cytoskeleton and accordingly affect subcellular localization. In conclusion, our data demonstrate that TRPM7 regulates myosin IIA filament stability and localization by phosphorylating a short stretch of amino acids within the alpha-helical tail of the myosin IIA heavy chain.
KW - Amino Acid Sequence
KW - Animals
KW - Cell Line
KW - Chlorocebus aethiops
KW - Conserved Sequence
KW - Humans
KW - Kinetics
KW - Mice
KW - Molecular Sequence Data
KW - Mutation/genetics
KW - Myosin Heavy Chains/metabolism
KW - Nonmuscle Myosin Type IIA/chemistry
KW - Phosphorylation
KW - Phosphoserine/metabolism
KW - Phosphothreonine/metabolism
KW - Sequence Alignment
KW - TRPM Cation Channels/genetics
U2 - 10.1016/j.jmb.2008.02.057
DO - 10.1016/j.jmb.2008.02.057
M3 - Article
C2 - 18394644
VL - 378
SP - 790
EP - 803
JO - Journal of molecular biology
JF - Journal of molecular biology
SN - 0022-2836
IS - 4
ER -