Redox-based control of the gamma heavy chain ATPase from Chlamydomonas outer arm dynein.
The outer dynein arm from Chlamydomonas flagella contains two redox-active thioredoxin-related light chains associated with the alpha and beta heavy chains; these proteins belong to a distinct subgroup within the thioredoxin family. This observation suggested that some aspect of dynein activity might be modulated through redox poise. To test this, we have examined the effect of sulfhydryl oxidation on the ATPase activity of isolated dynein and axonemes from wildtype and mutant strains lacking various heavy chain combinations. The outer, but not inner, dynein arm ATPase was stimulated significantly following treatment with low concentrations of dithionitrobenzoic acid; this effect was readily reversible by dithiol, and to a lesser extent, monothiol reductants. Mutational and biochemical dissection of the outer arm revealed that ATPase activation in response to DTNB was an exclusive property of the gamma heavy chain, and that enzymatic enhancement was modulated by the presence of other dynein components. Furthermore, we demonstrate that the LC5 thioredoxin-like light chain binds to the N-terminal stem domain of the alpha heavy chain and that the beta heavy chain-associated LC3 protein also interacts with the gamma heavy chain. These data suggest the possibility of a dynein-associated redox cascade and further support the idea that the gamma heavy chain plays a key regulatory role within the outer arm.
Department of Biochemistry, University of Connecticut Health Center, Farmington, Connecticut 06030-3305, USA.
Alistair
A
Harrison
Miho
M
Sakato
Hugo W
HW
Tedford
Sharon E
SE
Benashski
Ramila S
RS
Patel-King
Stephen M
SM
King
D000595:Amino Acid Sequence; D000818:Animals; D002696:Chlamydomonas / Q000201:enzymology*; D004228:Dithionitrobenzoic Acid / Q000378:metabolism; D004398:Dyneins / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D013329:Structure-Activity Relationship
D004228:Dithionitrobenzoic Acid; D004398:Dyneins