Genetic suppression of a phosphomimic myosin II identifies system-level factors that promote myosin II cleavage furrow accumulation

Yixin Ren, Hoku West-Foylea, Alexandra Surcel, Christopher Millera, Douglas N. Robinsona

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)4150-4165
Number of pages16
JournalMolecular Biology of the Cell
Volume25
Issue number25
DOIs
StatePublished - Dec 15 2014
Externally publishedYes

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Genetic Suppression
Myosin Type II
Cytokinesis
14-3-3 Proteins
Mechanical Stress
Dictyostelium
Genetic Selection
Myosins
Gene Library
Actins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Genetic suppression of a phosphomimic myosin II identifies system-level factors that promote myosin II cleavage furrow accumulation. / Ren, Yixin; West-Foylea, Hoku; Surcel, Alexandra; Millera, Christopher; Robinsona, Douglas N.

In: Molecular Biology of the Cell, Vol. 25, No. 25, 15.12.2014, p. 4150-4165.

Research output: Contribution to journalArticle

Ren, Yixin ; West-Foylea, Hoku ; Surcel, Alexandra ; Millera, Christopher ; Robinsona, Douglas N. / Genetic suppression of a phosphomimic myosin II identifies system-level factors that promote myosin II cleavage furrow accumulation. In: Molecular Biology of the Cell. 2014 ; Vol. 25, No. 25. pp. 4150-4165.
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