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. 1997 Sep;147(1):87–100. doi: 10.1093/genetics/147.1.87

In Vivo Analysis of Saccharomyces Cerevisiae Cox2 mRNA 5'-Untranslated Leader Functions in Mitochondrial Translation Initiation and Translational Activation

H M Dunstan 1, N S Green-Willms 1, T D Fox 1
PMCID: PMC1208125  PMID: 9286670

Abstract

We have used mutational and revertant analysis to study the elements of the 54-nucleotide COX2 5'-untranslated leader involved in translation initiation in yeast mitochondria and in activation by the COX2 translational activator, Pet111p. We generated a collection of mutants with substitutions spanning the entire COX2 5'-UTL by in vitro mutagenesis followed by mitochondrial transformation and gene replacement. The phenotypes of these mutants delimit a 31-nucleotide segment, from -16 to -46, that contains several short sequence elements necessary for COX2 5'-UTL function in translation. The sequences from -16 to -47 were shown to be partially sufficient to promote translation in a foreign context. Analysis of revertants of both the series of linker-scanning alleles and two short deletion/insertion alleles has refined the positions of several possible functional elements of the COX2 5'-untranslated leader, including a putative RNA stem-loop structure that functionally interacts with Pet111p and an octanucleotide sequence present in all S. cerevisiae mitochondrial mRNA 5'-UTLs that is a potential rRNA binding site.

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Selected References

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