Characterization of cytoplasmic factors which complement ca2 + channel mutations in paramecium tetraurelia

Nobuyuki Haga, Michael Forte, Yoshiro Saimi, Ching Kung

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The analysis of Ca2 +-channel function in the single-celled eukaryote Paramecium can be extended to a biochemical level based on recent observations that transfer of cytoplasm from wild-type cells into mutants lacking Ca2 + -channel function ("pawn" mutants) causes the mutant cells to regain Ca2 + -channel activity. Using a convenient behavioral assay for Ca2 + -channel function, we have used microinjection of cytoplasmic fractions into mutant cells to enrich for and characterize those components from wild-type cytoplasm which can "cure" cells carrying mutations in the 3 different pawn genes affecting Ca2+ -channel activity (pwA,pwB, and pwC). In each case, the curing factor appears to be a protein component of an intracellular membrane. They are distinguishable on the basis of thermal, pH and divalent ion sensitivities. In addition, the factor curing IhepwC mutational defect has been purified more than 180-fold. Furthermore, thepwB curing activity appears to be amplified during sequential transfer between pwB cells.

Original languageEnglish (US)
Pages (from-to)259-274
Number of pages16
JournalJournal of Neurogenetics
Volume1
Issue number3
DOIs
StatePublished - 1984
Externally publishedYes

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Paramecium tetraurelia
Mutation
Cytoplasm
Paramecium
Intracellular Membranes
Microinjections
Eukaryota
Hot Temperature
Ions

Keywords

  • Behavioral mutants
  • Microinjection
  • Paramecium tetraurelia

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Genetics
  • Developmental Biology
  • Neuroscience(all)

Cite this

Characterization of cytoplasmic factors which complement ca2 + channel mutations in paramecium tetraurelia. / Haga, Nobuyuki; Forte, Michael; Saimi, Yoshiro; Kung, Ching.

In: Journal of Neurogenetics, Vol. 1, No. 3, 1984, p. 259-274.

Research output: Contribution to journalArticle

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AU - Saimi, Yoshiro

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N2 - The analysis of Ca2 +-channel function in the single-celled eukaryote Paramecium can be extended to a biochemical level based on recent observations that transfer of cytoplasm from wild-type cells into mutants lacking Ca2 + -channel function ("pawn" mutants) causes the mutant cells to regain Ca2 + -channel activity. Using a convenient behavioral assay for Ca2 + -channel function, we have used microinjection of cytoplasmic fractions into mutant cells to enrich for and characterize those components from wild-type cytoplasm which can "cure" cells carrying mutations in the 3 different pawn genes affecting Ca2+ -channel activity (pwA,pwB, and pwC). In each case, the curing factor appears to be a protein component of an intracellular membrane. They are distinguishable on the basis of thermal, pH and divalent ion sensitivities. In addition, the factor curing IhepwC mutational defect has been purified more than 180-fold. Furthermore, thepwB curing activity appears to be amplified during sequential transfer between pwB cells.

AB - The analysis of Ca2 +-channel function in the single-celled eukaryote Paramecium can be extended to a biochemical level based on recent observations that transfer of cytoplasm from wild-type cells into mutants lacking Ca2 + -channel function ("pawn" mutants) causes the mutant cells to regain Ca2 + -channel activity. Using a convenient behavioral assay for Ca2 + -channel function, we have used microinjection of cytoplasmic fractions into mutant cells to enrich for and characterize those components from wild-type cytoplasm which can "cure" cells carrying mutations in the 3 different pawn genes affecting Ca2+ -channel activity (pwA,pwB, and pwC). In each case, the curing factor appears to be a protein component of an intracellular membrane. They are distinguishable on the basis of thermal, pH and divalent ion sensitivities. In addition, the factor curing IhepwC mutational defect has been purified more than 180-fold. Furthermore, thepwB curing activity appears to be amplified during sequential transfer between pwB cells.

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