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PMID:15987738

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Citation

Yumura, S, Yoshida, M, Betapudi, V, Licate, LS, Iwadate, Y, Nagasaki, A, Uyeda, TQ and Egelhoff, TT (2005) Multiple myosin II heavy chain kinases: roles in filament assembly control and proper cytokinesis in Dictyostelium. Mol. Biol. Cell 16:4256-66

Abstract

Myosin II filament assembly in Dictyostelium discoideum is regulated via phosphorylation of residues located in the carboxyl-terminal portion of the myosin II heavy chain (MHC) tail. A series of novel protein kinases in this system are capable of phosphorylating these residues in vitro, driving filament disassembly. Previous studies have demonstrated that at least three of these kinases (MHCK A, MHCK B, and MHCK C) display differential localization patterns in living cells. We have created a collection of single, double, and triple gene knockout cell lines for this family of kinases. Analysis of these lines reveals that three MHC kinases appear to represent the majority of cellular activity capable of driving myosin II filament disassembly, and reveals that cytokinesis defects increase with the number of kinases disrupted. Using biochemical fractionation of cytoskeletons and in vivo measurements via fluorescence recovery after photobleaching (FRAP), we find that myosin II overassembly increases incrementally in the mutants, with the MHCK A(-)/B(-)/C(-) triple mutant showing severe myosin II overassembly. These studies suggest that the full complement of MHC kinases that significantly contribute to growth phase and cytokinesis myosin II disassembly in this organism has now been identified.

Links

PubMed PMC1196335 Online version:10.1091/mbc.E05-03-0219

Keywords

Animals; Calcium-Calmodulin-Dependent Protein Kinases/genetics; Calcium-Calmodulin-Dependent Protein Kinases/physiology; Cell Line; Cytokinesis/physiology; Cytoskeleton/enzymology; Dictyostelium/enzymology; Dictyostelium/growth & development; Genes, Reporter; Myosin Heavy Chains/metabolism; Myosin Type II/genetics; Myosin Type II/metabolism; Myosin Type II/physiology; Phosphorylation; Protein Processing, Post-Translational; Protozoan Proteins; Recombinant Fusion Proteins/genetics; Recombinant Fusion Proteins/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DICDI:MHCKA

acts_upstream_of_or_within

GO:0000281: mitotic cytokinesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:MHCKA

acts_upstream_of_or_within

GO:0031037: myosin II filament disassembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:MHCKB

acts_upstream_of_or_within

GO:0000281: mitotic cytokinesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:MHCKB

located_in

GO:0005826: actomyosin contractile ring

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:MHCKB

located_in

GO:0005737: cytoplasm

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:MHCKB

acts_upstream_of_or_within

GO:0031037: myosin II filament disassembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:AK1

enables

GO:0005524: ATP binding

ECO:0000305: curator inference used in manual assertion

GO:0004674

F

Seeded From UniProt

complete

DICDI:AK1

acts_upstream_of_or_within

GO:0006468: protein phosphorylation

ECO:0000250: sequence similarity evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:AK1

enables

GO:0004674: protein serine/threonine kinase activity

ECO:0000250: sequence similarity evidence used in manual assertion

F

Seeded From UniProt

complete

DICDI:MHCKD

acts_upstream_of_or_within

GO:0031037: myosin II filament disassembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:MHCKC

acts_upstream_of_or_within

GO:0000281: mitotic cytokinesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DICDI:MHCKC

located_in

GO:0005826: actomyosin contractile ring

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DICDI:MHCKC

acts_upstream_of_or_within

GO:0031037: myosin II filament disassembly

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

See also

References

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