The presequence-binding domain of Tim50 is essential. (A) Diploid yeast cells carrying a single chromosomal deletion of TIM50366-476 were sporulated and subjected to tetrad dissection. (B) Yeast cells containing a chromosomal deletion of TIM50, complemented by a TIM50-containing plasmid carrying URA3, were transformed with plasmid constructs encoding the indicated Tim50 versions and subjected to plasmid loss on 5-FOA–containing medium. (C) Western blot analysis of mitochondria isolated from the cells described in A with the indicated antibodies. (D) In organello photo cross-linking of presequence probes using mitochondria from indicated strains for 30 min. Samples were analyzed as in C.
Articles from The Journal of Cell Biology are provided here courtesy of
The Rockefeller University Press