A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis.

E. André, M. Brink, G. Gerisch, G. Isenberg, A. Noegel, M. Schleicher, Jeffrey E. Segall, E. Wallraff

Research output: Contribution to journalArticle

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Abstract

A severin deficient mutant of Dictyostelium discoideum has been isolated by the use of colony immunoblotting after chemical mutagenesis. In homogenates of wild-type cells, severin is easily detected as a very active F-actin fragmenting protein. Tests for severin in the mutant, HG1132, included viscometry for the assay of F-actin fragmentation in fractions from DEAE-cellulose columns, labeling of blots with monoclonal and polyclonal antibodies, and immunofluorescent-labeling of cryosections. Severin could not be detected in the mutant using these methods. The mutation in HG1132 is recessive and has been mapped to linkage group VII. The mutant failed to produce the normal severin mRNA, but small amounts of a transcript that was approximately 100 bases larger than the wild-type mRNA were detected in the mutant throughout all stages of development. On the DNA level a new Mbo II restriction site was found in the mutant within the coding region of the severin gene. The severin deficient mutant cells grew at an approximately normal rate, aggregated and formed fruiting bodies with viable spores. By the use of an image processing system, speed of cell movement, turning rates, and precision of chemotactic orientation in a stable gradient of cyclic AMP were quantitated, and no significant differences between wild-type and mutant cells were found. Thus, under the culture conditions used, severin proved to be neither essential for growth of D. discoideum nor for any cell function that is important for aggregation or later development.

Original languageEnglish (US)
Pages (from-to)985-995
Number of pages11
JournalJournal of Cell Biology
Volume108
Issue number3
StatePublished - Mar 1989
Externally publishedYes

Fingerprint

Dictyostelium
Chemotaxis
Actins
Proteins
DEAE-Cellulose
Messenger RNA
Dictyostelium severin protein
Spores
Immunoblotting
Mutagenesis
Cyclic AMP
Cell Movement
Monoclonal Antibodies
Mutation
DNA
Growth

ASJC Scopus subject areas

  • Cell Biology

Cite this

André, E., Brink, M., Gerisch, G., Isenberg, G., Noegel, A., Schleicher, M., ... Wallraff, E. (1989). A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. Journal of Cell Biology, 108(3), 985-995.

A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. / André, E.; Brink, M.; Gerisch, G.; Isenberg, G.; Noegel, A.; Schleicher, M.; Segall, Jeffrey E.; Wallraff, E.

In: Journal of Cell Biology, Vol. 108, No. 3, 03.1989, p. 985-995.

Research output: Contribution to journalArticle

André, E, Brink, M, Gerisch, G, Isenberg, G, Noegel, A, Schleicher, M, Segall, JE & Wallraff, E 1989, 'A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis.', Journal of Cell Biology, vol. 108, no. 3, pp. 985-995.
André E, Brink M, Gerisch G, Isenberg G, Noegel A, Schleicher M et al. A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. Journal of Cell Biology. 1989 Mar;108(3):985-995.
André, E. ; Brink, M. ; Gerisch, G. ; Isenberg, G. ; Noegel, A. ; Schleicher, M. ; Segall, Jeffrey E. ; Wallraff, E. / A Dictyostelium mutant deficient in severin, an F-actin fragmenting protein, shows normal motility and chemotaxis. In: Journal of Cell Biology. 1989 ; Vol. 108, No. 3. pp. 985-995.
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abstract = "A severin deficient mutant of Dictyostelium discoideum has been isolated by the use of colony immunoblotting after chemical mutagenesis. In homogenates of wild-type cells, severin is easily detected as a very active F-actin fragmenting protein. Tests for severin in the mutant, HG1132, included viscometry for the assay of F-actin fragmentation in fractions from DEAE-cellulose columns, labeling of blots with monoclonal and polyclonal antibodies, and immunofluorescent-labeling of cryosections. Severin could not be detected in the mutant using these methods. The mutation in HG1132 is recessive and has been mapped to linkage group VII. The mutant failed to produce the normal severin mRNA, but small amounts of a transcript that was approximately 100 bases larger than the wild-type mRNA were detected in the mutant throughout all stages of development. On the DNA level a new Mbo II restriction site was found in the mutant within the coding region of the severin gene. The severin deficient mutant cells grew at an approximately normal rate, aggregated and formed fruiting bodies with viable spores. By the use of an image processing system, speed of cell movement, turning rates, and precision of chemotactic orientation in a stable gradient of cyclic AMP were quantitated, and no significant differences between wild-type and mutant cells were found. Thus, under the culture conditions used, severin proved to be neither essential for growth of D. discoideum nor for any cell function that is important for aggregation or later development.",
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