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Related Experiment Videos

Reversion analysis of dynein intermediate chain function

D R Mitchell1, Y Kang

  • 1Department of Anatomy and Cell Biology, SUNY Health Science Center, Syracuse 13210.

Journal of Cell Science
|August 1, 1993
PubMed
Summary
This summary is machine-generated.

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The ODA6 gene

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • The ODA6 locus in Chlamydomonas reinhardtii is crucial for flagellar outer row dynein assembly.
  • Mutations in ODA6 disrupt the entire outer row dynein arm complex formation.
  • Understanding the structure-function of the 70 kDa intermediate chain is key to flagellar motility.

Purpose of the Study:

  • To perform a structure-function analysis of the 70 kDa intermediate chain protein encoded by ODA6.
  • To investigate the role of the ODA6 intermediate chain in outer dynein arm assembly and function.
  • To elucidate the relationship between ODA6 mutations, protein structure, and flagellar beat frequency.

Main Methods:

  • Site-directed mutagenesis and gene transformation using chimeric mutant/wild-type ODA6 genes.

Related Experiment Videos

  • Selection and analysis of intragenic pseudorevertants of the oda6-95 mutation.
  • DNA sequencing, immunoblot analysis, and electron microscopy to characterize revertant alleles and protein assembly.

    • Main Results:

    • A frame-shift mutation in oda6-95 leads to a non-functional outer row dynein arm.
    • Pseudorevertants restored outer row arm assembly but exhibited varying flagellar beat frequencies (54 Hz or 27 Hz).
    • Motility defects in revertants did not correlate with deficits in specific dynein subunit assembly or outer arm structure.

    Conclusions:

    • The 70 kDa intermediate chain of outer dynein arms plays a direct role in flagellar function beyond its role in assembly.
    • The position and direction of secondary frame-shift mutations influence the severity of motility defects.
    • Structural alterations in the ODA6 intermediate chain directly impact flagellar motor performance.