TY - JOUR
T1 - Genetic suppression of a phosphomimic myosin II identifies system-level factors that promote myosin II cleavage furrow accumulation
AU - Ren, Yixin
AU - West-Foylea, Hoku
AU - Surcela, Alexra
AU - Millera, Christopher
AU - Robinsona, Douglas N.
N1 - Publisher Copyright:
© 2014 Ren et al. We thank the members of the Robinson lab and Pablo Iglesias for helpful discussions and comments on the manuscript. This work was supported by National Institutes of Health Grant GM066817 and support to the Summer Academic Research Experience program.
PY - 2014/12/15
Y1 - 2014/12/15
N2 - How myosin II localizes to the cleavage furrow in Dictyostelium and metazoan cells remains largely unknown despite significant advances in understanding its regulation. We designed a genetic selection using cDNA library suppression of 3×Asp myosin II to iden?tify factors involved in myosin cleavage furrow accumulation. The 3×Asp mutant is deficient in bipolar thick filament assembly, fails to accumulate at the cleavage furrow, cannot rescue myoII-null cytokinesis, and has impaired mechanosensitive accumulation. Eleven genes sup? pressed this dominant cytokinesis deficiency when 3×Asp was expressed in wild-type cells. 3×Asp myosin II's localization to the cleavage furrow was rescued by constructs encoding rcdBB, mmsdh, RMD1, actin, one novel protein, and a 14-3-3 hairpin. Further characterization showed that RMD1 is required for myosin II cleavage furrow accumulation, acting in parallel with mechanical stress. Analysis of several mutant strains revealed that different thresholds of myosin II activity are required for daughter cell symmetry than for furrow ingression dy? namics. Finally, an engineered myosin II with a longer lever arm (2×ELC), producing a highly mechanosensitive motor, could also partially suppress the intragenic 3×Asp. Overall, myosin II accumulation is the result of multiple parallel and partially redundant pathways that com? prise a cellular contractility control system.
AB - How myosin II localizes to the cleavage furrow in Dictyostelium and metazoan cells remains largely unknown despite significant advances in understanding its regulation. We designed a genetic selection using cDNA library suppression of 3×Asp myosin II to iden?tify factors involved in myosin cleavage furrow accumulation. The 3×Asp mutant is deficient in bipolar thick filament assembly, fails to accumulate at the cleavage furrow, cannot rescue myoII-null cytokinesis, and has impaired mechanosensitive accumulation. Eleven genes sup? pressed this dominant cytokinesis deficiency when 3×Asp was expressed in wild-type cells. 3×Asp myosin II's localization to the cleavage furrow was rescued by constructs encoding rcdBB, mmsdh, RMD1, actin, one novel protein, and a 14-3-3 hairpin. Further characterization showed that RMD1 is required for myosin II cleavage furrow accumulation, acting in parallel with mechanical stress. Analysis of several mutant strains revealed that different thresholds of myosin II activity are required for daughter cell symmetry than for furrow ingression dy? namics. Finally, an engineered myosin II with a longer lever arm (2×ELC), producing a highly mechanosensitive motor, could also partially suppress the intragenic 3×Asp. Overall, myosin II accumulation is the result of multiple parallel and partially redundant pathways that com? prise a cellular contractility control system.
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U2 - 10.1091/mbc.E14-08-1322
DO - 10.1091/mbc.E14-08-1322
M3 - Article
C2 - 25318674
AN - SCOPUS:84919478111
SN - 1059-1524
VL - 25
SP - 4150
EP - 4165
JO - Molecular biology of the cell
JF - Molecular biology of the cell
IS - 25
ER -