Abstract
Coupling factor slow recovery (cfs) is a recessive mutant of Arabidopsis with anomalous ATP synthase activation/deactivation characteristics as well as a distinct growth phenotype. The most significant feature of this mutant is that the dark-adapted deactivation of ATP synthase is a very slow relative to the wild type, indicating interference with ATP synthase regulation. Physical mapping of the mutation delimited it to a region in a pair of bacterial artificial chromosome clones. Examination of T-DNA insertion lines of all 34 putative genes located in this region identified two homozygous T-DNA insertion lines of the same gene, At3g59770, possessing phenotypes indistinguishable from the cfs mutant. At3g59770 had been previously identified as suppressor of actin 9 (SAC9), a protein with a SAC domain, a protein–protein interaction module containing two conserved tryptophans known as a WW domain, and an ATP/GTP-binding site motif A. Sequence analysis of cfs revealed a point mutation of G to A resulting in an amino acid substitution from tryptophan to STOP, thereby coding a truncated protein. Real-time-PCR amplification of the gene specific fragments showed that the T-DNA mutants did not have full-length transcripts whereas the cfs mutant transcribed a full-length mutated transcript. Further investigation of SAC9 RNA expression levels in different tissues of wild-type plants by RT-PCR revealed the highest expression in leaves. SAC 9 dysfunction interferes with ATP synthase deactivation, possibly by an alteration in phosphoinositide signaling inducing a stress mimicry response.
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Abbreviations
- EMS:
-
ethylmethanesulfonate
- cfs :
-
coupling factor slow recovery
- SAC:
-
suppressor of actin
- T-DNA:
-
transfer DNA
- Δμ +H :
-
protonmotive force
- CAPS:
-
cleaved amplified polymorphic sequences
- SSLP:
-
simple sequence length polymorphisms
- MMLV:
-
Maloney murine leukemia virus
- DCCD:
-
N,N′-dicyclohexylcarbodiimide
- MS medium:
-
Murashige and Skoog medium
- ER:
-
endoplasmic reticulum
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Acknowledgements
We are grateful to Dr. Aleel Grennan for her insightful comments and expert editorial assistance. We also thank Dr. Mary Williams for her valuable comments and for sharing her ideas with us.
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Gong, P., Wu, G. & Ort, D.R. Slow dark deactivation of Arabidopsis chloroplast ATP synthase caused by a mutation in a nonplastidic SAC domain protein. Photosynth Res 88, 133–142 (2006). https://doi.org/10.1007/s11120-006-9041-4
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DOI: https://doi.org/10.1007/s11120-006-9041-4