First published online March 6, 2003; 10.1104/pp.102.016808
Plant Physiol, April 2003, Vol. 131, pp. 1737-1747
Identification, Expression, and Import of Components 17 and
23 of the Inner Mitochondrial Membrane Translocase from
Arabidopsis1,[w]
Monika W.
Murcha,
Ryan
Lister,
Angela Y. Y.
Ho, and
James
Whelan*
Plant Molecular Biology Group, Biochemistry and Molecular Biology,
School of Biomedical and Chemical Sciences (M.W.M., R.L., A.Y.Y.H.,
J.W.) and Plant Biology, School of Natural and Agricultural Sciences
(M.W.M.), University of Western Australia, 35 Stirling Highway, Crawley
6009, Western Australia, Australia
Characterization of components 17 and 23 of the inner mitochondrial
membrane translocase (TIM17:23) from Arabidopsis indicated that there
were three genes present for TIM17 and
TIM23 and two for TIM44.
AtTIM17 differed from the yeast (Saccharomyces
cerevisiae) and mammalian homologs in that two genes encoded
proteins that were longer and one gene encoded a shorter protein. All
Arabidopsis TIM23 predicted proteins appeared to lack the first 34 amino acids compared with yeast TIM23. All AtTIM17 and
AtTIM23 genes were expressed but displayed different
tissue and developmental profiles. Complementation of deletion mutants
in yeast indicated that for AtTIM17, the extension at
the C terminus not present in yeast had to be removed to achieve
complementation, whereas for TIM23, a preprotein and
amino acid transporter domain had to be present for complementation.
Import assays with AtTIM17 and AtTIM23
indicated that they both contained internal signals for integration
into the inner mitochondrial membrane in a membrane potential-dependent manner. The C terminus of imported AtTIM17-2 was
susceptible to degradation by externally added protease with intact
mitochondria. Removal of the 85 C-terminal amino acids resulted in
import and full protection of the truncated protein. This suggests that
the novel extension at the C terminus of AtTIM17-2 links
the outer and inner membrane in a manner analogous to yeast TIM23.
1
This work was supported by the Australian
Research Council (grant to J.W.).
[w]
The online version of this article contains Web-only
data. The supplemental material is available at
www.plantphysiol.org.
*
Corresponding author; e-mail seamus{at}cyllene.uwa.edu.au; fax
61-8-93801148.
© 2003 American Society of Plant Biologists
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