We have developed a system for performing interaction genetics in Dictyostelium discoideum that uses a cDNA library complementation/multicopy suppression strategy. Chemically mutagenized cells were screened for cytokinesis-deficient mutants and one mutant was subjected to library complementation. Isolates of four different genes were recovered as modifiers of this strain's cytokinesis defect. These include the cleavage furrow protein cortexillin I, a novel protein we named dynacortin, an ezrin-radixin-moesin-family protein, and coronin. The cortexillin I locus and transcript were found to be disrupted in the strain, identifying it as the affected gene. Dynacortin is localized partly to the cell cortex and becomes enriched in protrusive regions, a localization pattern that is similar to coronin and partly dependent on RacE. During cytokinesis, dynacortin is found in the cortex and is somewhat enriched at the poles. Furthermore, it appears to be reduced in the cleavage furrow. The genetic interactions and the cellular distributions of the proteins suggest a hypothesis for cytokinesis in which the contraction of the medial ring is a function of spatially restricted cortexillin I and myosin II and globally distributed dynacortin, coronin, and RacE.
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21 August 2000
Article|
August 21 2000
Dynacortin, a Genetic Link between Equatorial Contractility and Global Shape Control Discovered by Library Complementation of a Dictyostelium discoideum Cytokinesis Mutant
Douglas N. Robinson,
Douglas N. Robinson
aDepartment of Biochemistry and Developmental Biology, Beckman Center, Stanford University, Stanford, California 94305-5307
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James A. Spudich
James A. Spudich
aDepartment of Biochemistry and Developmental Biology, Beckman Center, Stanford University, Stanford, California 94305-5307
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Douglas N. Robinson
aDepartment of Biochemistry and Developmental Biology, Beckman Center, Stanford University, Stanford, California 94305-5307
James A. Spudich
aDepartment of Biochemistry and Developmental Biology, Beckman Center, Stanford University, Stanford, California 94305-5307
Abbreviations used in this paper: myo II, myosin II; myo II−, myo II null cell line; NQNO, 4-nitroquinolone-N-oxide; NaVi, sodium vanadate (Na2HVO4); ORF, open reading frame.
Received:
March 07 2000
Revision Requested:
June 20 2000
Accepted:
June 28 2000
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Cell Biol (2000) 150 (4): 823–838.
Article history
Received:
March 07 2000
Revision Requested:
June 20 2000
Accepted:
June 28 2000
Citation
Douglas N. Robinson, James A. Spudich; Dynacortin, a Genetic Link between Equatorial Contractility and Global Shape Control Discovered by Library Complementation of a Dictyostelium discoideum Cytokinesis Mutant. J Cell Biol 21 August 2000; 150 (4): 823–838. doi: https://doi.org/10.1083/jcb.150.4.823
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